published_papers
"タイトル(日本語)","タイトル(英語)","著者(日本語)","著者(英語)","担当区分","概要(日本語)","概要(英語)","出版者・発行元(日本語)","出版者・発行元(英語)","出版年月","誌名(日本語)","誌名(英語)","巻","号","開始ページ","終了ページ","記述言語","査読の有無","招待の有無","掲載種別","国際・国内誌","国際共著","DOI","ISSN","eISSN","URL","URL2","主要な業績かどうか","公開の有無"
"Protective effect of oleic acid against very long-chain fatty acid-induced apoptosis in peroxisome-deficient CHO cells","Protective effect of oleic acid against very long-chain fatty acid-induced apoptosis in peroxisome-deficient CHO cells","Hanif ALi, Mone Yamanishi, Keigo Sunagawa, Mizuki Kumon, Rumana Yesmin Hasi, Mutsumi Aihara, Ryushi Kawakami, Tamotsu Tanaka","Hanif ALi, Mone Yamanishi, Keigo Sunagawa, Mizuki Kumon, Rumana Yesmin Hasi, Mutsumi Aihara, Ryushi Kawakami, Tamotsu Tanaka","null","Very long-chain fatty acids (VLCFAs) are degraded exclusively in peroxisomes, as evidenced by the accumulation of VLCFAs in patients with certain peroxisomal disorders. Although accumulation of VLCFAs is considered to be associated with health issues, including neuronal degeneration, the mechanisms underlying VLCFAs-induced tissue degeneration remain unclear. Here, we report the toxic effect of VLCFA and protective effect of C18: 1 FA in peroxisome-deficient CHO cells. We examined the cytotoxicity of saturated and monounsaturated VLCFAs with chain-length at C20-C26, and found that longer and saturated VLCFA showed potent cytotoxicity at lower accumulation levels. Furthermore, the extent of VLCFA-induced toxicity was found to be associated with a decrease in cellular C18:1 FA levels. Notably, supplementation with C18:1 FA effectively rescued the cells from VLCFA-induced apoptosis without reducing the cellular VLCFAs levels, implying that peroxisome-deficient cells can survive in the presence of accumulated VLCFA, as long as the cells keep sufficient levels of cellular C18:1 FA. These results suggest a therapeutic potential of C18:1 FA in peroxisome disease and may provide new insights into the pharmacological effect of Lorenzo's oil, a 4:1 mixture of C18:1 and C22:1 FA.","Very long-chain fatty acids (VLCFAs) are degraded exclusively in peroxisomes, as evidenced by the accumulation of VLCFAs in patients with certain peroxisomal disorders. Although accumulation of VLCFAs is considered to be associated with health issues, including neuronal degeneration, the mechanisms underlying VLCFAs-induced tissue degeneration remain unclear. Here, we report the toxic effect of VLCFA and protective effect of C18: 1 FA in peroxisome-deficient CHO cells. We examined the cytotoxicity of saturated and monounsaturated VLCFAs with chain-length at C20-C26, and found that longer and saturated VLCFA showed potent cytotoxicity at lower accumulation levels. Furthermore, the extent of VLCFA-induced toxicity was found to be associated with a decrease in cellular C18:1 FA levels. Notably, supplementation with C18:1 FA effectively rescued the cells from VLCFA-induced apoptosis without reducing the cellular VLCFAs levels, implying that peroxisome-deficient cells can survive in the presence of accumulated VLCFA, as long as the cells keep sufficient levels of cellular C18:1 FA. These results suggest a therapeutic potential of C18:1 FA in peroxisome disease and may provide new insights into the pharmacological effect of Lorenzo's oil, a 4:1 mixture of C18:1 and C22:1 FA.","null","null","2024-01-18","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1869","No.3","159452","159452","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2024.159452","1879-2618","null","null","null","null","null"
"Degradation of glycosylinositol phosphoceramide during plant tissue homogenization","Degradation of glycosylinositol phosphoceramide during plant tissue homogenization","Yoshimichi Takai, Rumana Yesmin Hasi, Naoko Matsumoto, Chiho Fujita, Hanif Ali, Junji Hayashi, Ryushi Kawakami, Mutsumi Aihara, Toshiki Ishikawa, Hiroyuki Imai, Mayuko Wakida, Kazuya Ando, Tamotsu Tanaka","Yoshimichi Takai, Rumana Yesmin Hasi, Naoko Matsumoto, Chiho Fujita, Hanif Ali, Junji Hayashi, Ryushi Kawakami, Mutsumi Aihara, Toshiki Ishikawa, Hiroyuki Imai, Mayuko Wakida, Kazuya Ando, Tamotsu Tanaka","null","null","null","null","null","2024-01","The Journal of Biochemistry","The Journal of Biochemistry","Vol.175","No.1","115","124","eng","true","null","scientific_journal","null","null","10.1093/jb/mvad080","1756-2651","null","null","null","null","null"
"Sphingosine kinase 1 is involved in triglyceride breakdown by maintaining lysosomal integrity in brown adipocytes","Sphingosine kinase 1 is involved in triglyceride breakdown by maintaining lysosomal integrity in brown adipocytes","Jun-ichi Morishige, Kazuaki Yoshioka, Hiroki Nakata, Kazuhiro Ishimaru, Naoto Nagata, Tamotsu Tanaka, Yoh Takuwa, Hitoshi Ando","Jun-ichi Morishige, Kazuaki Yoshioka, Hiroki Nakata, Kazuhiro Ishimaru, Naoto Nagata, Tamotsu Tanaka, Yoh Takuwa, Hitoshi Ando","null","Sphingosine 1-phosphate (S1P) has been implicated in brown adipose tissue (BAT) formation and energy consumption; however, the mechanistic role of sphingolipids, including S1P, in BAT remains unclear. Here, we showed that, in mice, BAT activation by cold exposure upregulated mRNA and protein expression of the S1P-synthesizing enzyme sphingosine kinase 1 (SphK1) and S1P production in BAT. Treatment of wild-type brown adipocytes with exogenous S1P or S1P receptor subtype-selective agonists stimulated triglyceride (TG) breakdown only marginally, compared with noradrenaline. However, genetic deletion of Sphk1 resulted in hypothermia and diminished body weight loss upon cold exposure, suggesting that SphK1 is involved in thermogenesis through mechanisms different from receptor-mediated, extracellular action of S1P. In BAT of wild-type mice, SphK1 was localized largely in the lysosomes of brown adipocytes. In the brown adipocytes of Sphk1 mice, the number of lysosomes was reduced and lysosomal function, including proteolytic activity, acid esterase activity, and motility, was impaired. Concordantly, nuclear translocation of transcription factor EB, a master transcriptional regulator of lysosome biogenesis, was reduced, leading to decreased mRNA expression of the lysosome-related genes in Sphk1 BAT. Moreover, BAT of Sphk1 mice showed greater TG accumulation with dominant larger lipid droplets in brown adipocytes. Inhibition of lysosomes with chloroquine resulted in a less extent of triglyceride accumulation in Sphk1 brown adipocytes compared with wild-type brown adipocytes, suggesting a reduced lysosome-mediated TG breakdown in Sphk1 mice. Our results indicate a novel role of SphK1 in lysosomal integrity, which is required for TG breakdown and thermogenesis in BAT.","Sphingosine 1-phosphate (S1P) has been implicated in brown adipose tissue (BAT) formation and energy consumption; however, the mechanistic role of sphingolipids, including S1P, in BAT remains unclear. Here, we showed that, in mice, BAT activation by cold exposure upregulated mRNA and protein expression of the S1P-synthesizing enzyme sphingosine kinase 1 (SphK1) and S1P production in BAT. Treatment of wild-type brown adipocytes with exogenous S1P or S1P receptor subtype-selective agonists stimulated triglyceride (TG) breakdown only marginally, compared with noradrenaline. However, genetic deletion of Sphk1 resulted in hypothermia and diminished body weight loss upon cold exposure, suggesting that SphK1 is involved in thermogenesis through mechanisms different from receptor-mediated, extracellular action of S1P. In BAT of wild-type mice, SphK1 was localized largely in the lysosomes of brown adipocytes. In the brown adipocytes of Sphk1 mice, the number of lysosomes was reduced and lysosomal function, including proteolytic activity, acid esterase activity, and motility, was impaired. Concordantly, nuclear translocation of transcription factor EB, a master transcriptional regulator of lysosome biogenesis, was reduced, leading to decreased mRNA expression of the lysosome-related genes in Sphk1 BAT. Moreover, BAT of Sphk1 mice showed greater TG accumulation with dominant larger lipid droplets in brown adipocytes. Inhibition of lysosomes with chloroquine resulted in a less extent of triglyceride accumulation in Sphk1 brown adipocytes compared with wild-type brown adipocytes, suggesting a reduced lysosome-mediated TG breakdown in Sphk1 mice. Our results indicate a novel role of SphK1 in lysosomal integrity, which is required for TG breakdown and thermogenesis in BAT.","null","null","2023-09-24","Journal of Lipid Research","Journal of Lipid Research","Vol.64","No.11","100450","100450","eng","true","null","scientific_journal","null","null","10.1016/j.jlr.2023.100450","1539-7262","null","https://doi.org/10.1016/j.jlr.2023.100450","null","null","null"
"GDE7 produces cyclic phsphpatidic acid in the ER lumen functioning as a lysophospholipid mediator","GDE7 produces cyclic phsphpatidic acid in the ER lumen functioning as a lysophospholipid mediator","Keisuke Kitakaza, Hanif Ali, Raiki Kimoto, Yasuhiro Takenouchi, Hironobu Ishimaru, Atsushi Yamashita, Natsuo Ueda, Tamotsu Tanaka, Yasuo Okamoto, Kazuhito Tsuboi","Keisuke Kitakaza, Hanif Ali, Raiki Kimoto, Yasuhiro Takenouchi, Hironobu Ishimaru, Atsushi Yamashita, Natsuo Ueda, Tamotsu Tanaka, Yasuo Okamoto, Kazuhito Tsuboi","null","Cyclic phosphatidic acid (cPA) is a lipid mediator, which regulates adipogenic differentiation and glucose homeostasis by suppressing nuclear peroxisome proliferator-activated receptor γ (PPARγ). Glycerophosphodiesterase 7 (GDE7) is a Ca-dependent lysophospholipase D that localizes in the endoplasmic reticulum. Although mouse GDE7 catalyzes cPA production in a cell-free system, it is unknown whether GDE7 generates cPA in living cells. Here, we demonstrate that human GDE7 possesses cPA-producing activity in living cells as well as in a cell-free system. Furthermore, the active site of human GDE7 is directed towards the luminal side of the endoplasmic reticulum. Mutagenesis revealed that amino acid residues F227 and Y238 are important for catalytic activity. GDE7 suppresses the PPARγ pathway in human mammary MCF-7 and mouse preadipocyte 3T3-L1 cells, suggesting that cPA functions as an intracellular lipid mediator. These findings lead to a better understanding of the biological role of GDE7 and its product, cPA.","Cyclic phosphatidic acid (cPA) is a lipid mediator, which regulates adipogenic differentiation and glucose homeostasis by suppressing nuclear peroxisome proliferator-activated receptor γ (PPARγ). Glycerophosphodiesterase 7 (GDE7) is a Ca-dependent lysophospholipase D that localizes in the endoplasmic reticulum. Although mouse GDE7 catalyzes cPA production in a cell-free system, it is unknown whether GDE7 generates cPA in living cells. Here, we demonstrate that human GDE7 possesses cPA-producing activity in living cells as well as in a cell-free system. Furthermore, the active site of human GDE7 is directed towards the luminal side of the endoplasmic reticulum. Mutagenesis revealed that amino acid residues F227 and Y238 are important for catalytic activity. GDE7 suppresses the PPARγ pathway in human mammary MCF-7 and mouse preadipocyte 3T3-L1 cells, suggesting that cPA functions as an intracellular lipid mediator. These findings lead to a better understanding of the biological role of GDE7 and its product, cPA.","null","null","2023-05-16","Communications Biology","Communications Biology","Vol.6","No.1","524","524","eng","true","null","scientific_journal","null","null","10.1038/s42003-023-04900-4","2399-3642","null","null","null","null","null"
"Molecular species profiles of plasma ceramides in different clinical types of X-linked adrenoleukodystrophy","Molecular species profiles of plasma ceramides in different clinical types of X-linked adrenoleukodystrophy","Morito Katsuya, Shimizu Ryota, Ali Hanif, Shimada Akina, Miyazaki Tohru, Takahashi Naoko, Rahman Motiur M., Tsuji Kazuki, Shimozawa Nobuyuki, Michiyasu Nakao, Shigeki Sano, Momoyo Azuma, Nanjundan Meera, Kentaro Kogure, Tamotsu Tanaka","Morito Katsuya, Shimizu Ryota, Ali Hanif, Shimada Akina, Miyazaki Tohru, Takahashi Naoko, Rahman Motiur M., Tsuji Kazuki, Shimozawa Nobuyuki, Michiyasu Nakao, Shigeki Sano, Momoyo Azuma, Nanjundan Meera, Kentaro Kogure, Tamotsu Tanaka","null","X-linked adrenoleukodystrophy (X-ALD) is a genetic disorder associated with peroxisomal dysfunction. Patients with this rare disease accumulate very long-chain fatty acids (VLCFAs) in their bodies because of impairment of peroxisomal VLCFA ?-oxidation. Several clinical types of X-ALD, ranging from mild (axonopathy in the spinal cord) to severe (cerebral demyelination), are known. However, the molecular basis for this phenotypic variability remains largely unknown. In this study, we determined plasma ceramide (CER) profile using liquid chromatography-tandem mass spectrometry. We characterized the molecular species profile of CER in the plasma of patients with mild (adrenomyeloneuropathy;AMN) and severe (cerebral) X-ALD. Eleven X-ALD patients (five cerebral, five AMN, and one carrier) and 10 healthy volunteers participated in this study. Elevation of C26:0 CER was found to be a common feature regardless of the clinical types. The level of C26:1 CER was significantly higher in AMN but not in cerebral type, than that in healthy controls. The C26:1 CER level in the cerebral type was significantly lower than that in the AMN type. These results suggest that a high level of C26:0 CER, along with a control level of C26:1 CER, is a characteristic feature of the cerebral type X-ALD. J. Med. Invest. 70 : 403-410, August, 2023.","X-linked adrenoleukodystrophy (X-ALD) is a genetic disorder associated with peroxisomal dysfunction. Patients with this rare disease accumulate very long-chain fatty acids (VLCFAs) in their bodies because of impairment of peroxisomal VLCFA ?-oxidation. Several clinical types of X-ALD, ranging from mild (axonopathy in the spinal cord) to severe (cerebral demyelination), are known. However, the molecular basis for this phenotypic variability remains largely unknown. In this study, we determined plasma ceramide (CER) profile using liquid chromatography-tandem mass spectrometry. We characterized the molecular species profile of CER in the plasma of patients with mild (adrenomyeloneuropathy;AMN) and severe (cerebral) X-ALD. Eleven X-ALD patients (five cerebral, five AMN, and one carrier) and 10 healthy volunteers participated in this study. Elevation of C26:0 CER was found to be a common feature regardless of the clinical types. The level of C26:1 CER was significantly higher in AMN but not in cerebral type, than that in healthy controls. The C26:1 CER level in the cerebral type was significantly lower than that in the AMN type. These results suggest that a high level of C26:0 CER, along with a control level of C26:1 CER, is a characteristic feature of the cerebral type X-ALD. J. Med. Invest. 70 : 403-410, August, 2023.","null","null","2023","The Journal of Medical Investigation : JMI","The Journal of Medical Investigation : JMI","Vol.70","No.3.4","403","410","eng","true","null","scientific_journal","null","null","10.2152/jmi.70.403","1349-6867","null","null","null","null","null"
"Peroxisomes attenuate cytotoxicity of very long-chain fatty acids","Peroxisomes attenuate cytotoxicity of very long-chain fatty acids","Hanif Ali, Miyu Kobayashi, Katsuya Morito, Rumana Yesmin, Mutsumi Aihara, Junji Hayashi, Ryushi Kawakami, Koichiro Tsuchiya, Kazunori Sango, Tamotsu Tanaka","Hanif Ali, Miyu Kobayashi, Katsuya Morito, Rumana Yesmin, Mutsumi Aihara, Junji Hayashi, Ryushi Kawakami, Koichiro Tsuchiya, Kazunori Sango, Tamotsu Tanaka","null","One of the major functions of peroxisomes in mammals is oxidation of very long-chain fatty acids (VLCFAs). Genetic defects in peroxisomal β-oxidation result in the accumulation of VLCFAs and lead to a variety of health problems, such as demyelination of nervous tissues. However, the mechanisms by which VLCFAs cause tissue degeneration have not been fully elucidated. Recently, we found that the addition of small amounts of isopropanol can enhance the solubility of saturated VLCFAs in an aqueous medium. In this study, we characterized the biological effect of extracellular VLCFAs in peroxisome-deficient Chinese hamster ovary (CHO) cells, neural crest-derived pheochromocytoma cells (PC12), and immortalized adult Fischer rat Schwann cells (IFRS1) using this solubilizing technique. C20:0 FA was the most toxic of the C16-C26 FAs tested in all cells. The basis of the toxicity of C20:0 FA was apoptosis and was observed at 5 μM and 30 μM in peroxisome-deficient and wild-type CHO cells, respectively. The sensitivity of wild-type CHO cells to cytotoxic C20:0 FA was enhanced in the presence of a peroxisomal β-oxidation inhibitor. Further, a positive correlation was evident between cell toxicity and the extent of intracellular accumulation of toxic FA. These results suggest that peroxisomes are pivotal in the detoxification of apoptotic VLCFAs by preventing their accumulation.","One of the major functions of peroxisomes in mammals is oxidation of very long-chain fatty acids (VLCFAs). Genetic defects in peroxisomal β-oxidation result in the accumulation of VLCFAs and lead to a variety of health problems, such as demyelination of nervous tissues. However, the mechanisms by which VLCFAs cause tissue degeneration have not been fully elucidated. Recently, we found that the addition of small amounts of isopropanol can enhance the solubility of saturated VLCFAs in an aqueous medium. In this study, we characterized the biological effect of extracellular VLCFAs in peroxisome-deficient Chinese hamster ovary (CHO) cells, neural crest-derived pheochromocytoma cells (PC12), and immortalized adult Fischer rat Schwann cells (IFRS1) using this solubilizing technique. C20:0 FA was the most toxic of the C16-C26 FAs tested in all cells. The basis of the toxicity of C20:0 FA was apoptosis and was observed at 5 μM and 30 μM in peroxisome-deficient and wild-type CHO cells, respectively. The sensitivity of wild-type CHO cells to cytotoxic C20:0 FA was enhanced in the presence of a peroxisomal β-oxidation inhibitor. Further, a positive correlation was evident between cell toxicity and the extent of intracellular accumulation of toxic FA. These results suggest that peroxisomes are pivotal in the detoxification of apoptotic VLCFAs by preventing their accumulation.","null","null","2023","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1868","No.2","159259","159259","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2022.159259","1879-2618","null","null","null","null","null"
"Nonspecific phospholipase C3 of radish has phospholipase D activity toward glycosylinositol phosphoceramide","Nonspecific phospholipase C3 of radish has phospholipase D activity toward glycosylinositol phosphoceramide","Rumana Yesmin Hasi, Toshiki Ishikawa, Keigo Sunagawa, Yoshimichi Takai, Hanif Ali, Junji Hayashi, Ryushi Kawakami, Keizo Yuasa, Mutsumi Aihara, Kaori Kanemaru, Hiroyuki Imai, Tamotsu Tanaka","Rumana Yesmin Hasi, Toshiki Ishikawa, Keigo Sunagawa, Yoshimichi Takai, Hanif Ali, Junji Hayashi, Ryushi Kawakami, Keizo Yuasa, Mutsumi Aihara, Kaori Kanemaru, Hiroyuki Imai, Tamotsu Tanaka","null","Glycosylinositol phosphoceramide (GIPC) is a major sphingolipid in the plasma membranes of plants. Previously, we found an enzyme activity that produces phytoceramide 1-phosphate (PC1P) by hydrolysis of the D position of GIPC in cabbage and named this activity as GIPC-phospholipase D (PLD). Here, we purified GIPC-PLD by sequential chromatography from radish roots. Peptide mass fingerprinting analysis revealed that the potential candidate for GIPC-PLD protein was nonspecific phospholipase C3 (NPC3), which has not been characterized as a PLD. The recombinant NPC3 protein obtained by heterologous expression system in Escherichia coli produced PC1P from GIPC and showed essentially the same enzymatic properties as those we characterized as GIPC-PLD in cabbage, radish and Arabidopsis thaliana. From these results, we conclude that NPC3 is one of the enzymes that degrade GIPC.","Glycosylinositol phosphoceramide (GIPC) is a major sphingolipid in the plasma membranes of plants. Previously, we found an enzyme activity that produces phytoceramide 1-phosphate (PC1P) by hydrolysis of the D position of GIPC in cabbage and named this activity as GIPC-phospholipase D (PLD). Here, we purified GIPC-PLD by sequential chromatography from radish roots. Peptide mass fingerprinting analysis revealed that the potential candidate for GIPC-PLD protein was nonspecific phospholipase C3 (NPC3), which has not been characterized as a PLD. The recombinant NPC3 protein obtained by heterologous expression system in Escherichia coli produced PC1P from GIPC and showed essentially the same enzymatic properties as those we characterized as GIPC-PLD in cabbage, radish and Arabidopsis thaliana. From these results, we conclude that NPC3 is one of the enzymes that degrade GIPC.","null","null","2022-10","FEBS Letters","FEBS Letters","Vol.596","No.23","3024","3036","eng","true","null","scientific_journal","null","null","10.1002/1873-3468.14520","1873-3468","null","null","null","null","null"
"Formation of N-acyl-phosphatidylethanolamines by cytosolic phospholipase A2ϵ in an ex vivo murine model of brain ischemia","Formation of N-acyl-phosphatidylethanolamines by cytosolic phospholipase A2ϵ in an ex vivo murine model of brain ischemia","S. Khaledur M. Rahman, Zahir Hussain, Katsuya Morito, Naoko Takahashi, Mohammad Mamun Sikder, Tamotsu Tanaka, Ken-ichi Ohta, Masaki Ueno, Hiroo Takahashi, Tohru Yamamoto, Makoto Murakami, Toru Uyama, Natsuo Ueda","S. Khaledur M. Rahman, Zahir Hussain, Katsuya Morito, Naoko Takahashi, Mohammad Mamun Sikder, Tamotsu Tanaka, Ken-ichi Ohta, Masaki Ueno, Hiroo Takahashi, Tohru Yamamoto, Makoto Murakami, Toru Uyama, Natsuo Ueda","null","N-Acyl-phosphatidylethanolamines (NAPEs), a minor class of membrane glycerophospholipids, accumulate along with their bioactive metabolites, N-acylethanolamines (NAEs) during ischemia. NAPEs can be formed through N-acylation of phosphatidylethanolamine by cytosolic phospholipase Aϵ (cPLAϵ, also known as PLA2G4E) or members of the phospholipase A and acyltransferase (PLAAT) family. However, the enzyme responsible for the NAPE production in brain ischemia has not yet been clarified. Here, we investigated a possible role of cPLAϵ using cPLAϵ-deficient (Pla2g4e) mice. As analyzed with brain homogenates of wild-type mice, the age dependency of Ca-dependent NAPE-forming activity showed a bell-shape pattern being the highest at the first week of postnatal life, and the activity was completely abolished in Pla2g4e mice. However, liquid chromatography-tandem mass spectrometry revealed that the NAPE levels of normal brain were similar between wild-type and Pla2g4e mice. In contrast, post-mortal accumulations of NAPEs and most species of NAEs were only observed in decapitated brains of wild-type mice. These results suggested that cPLAϵ is responsible for Ca-dependent formation of NAPEs in the brain as well as the accumulation of NAPEs and NAEs during ischemia, while other enzyme(s) appeared to be involved in the maintenance of basal NAPE levels.","N-Acyl-phosphatidylethanolamines (NAPEs), a minor class of membrane glycerophospholipids, accumulate along with their bioactive metabolites, N-acylethanolamines (NAEs) during ischemia. NAPEs can be formed through N-acylation of phosphatidylethanolamine by cytosolic phospholipase Aϵ (cPLAϵ, also known as PLA2G4E) or members of the phospholipase A and acyltransferase (PLAAT) family. However, the enzyme responsible for the NAPE production in brain ischemia has not yet been clarified. Here, we investigated a possible role of cPLAϵ using cPLAϵ-deficient (Pla2g4e) mice. As analyzed with brain homogenates of wild-type mice, the age dependency of Ca-dependent NAPE-forming activity showed a bell-shape pattern being the highest at the first week of postnatal life, and the activity was completely abolished in Pla2g4e mice. However, liquid chromatography-tandem mass spectrometry revealed that the NAPE levels of normal brain were similar between wild-type and Pla2g4e mice. In contrast, post-mortal accumulations of NAPEs and most species of NAEs were only observed in decapitated brains of wild-type mice. These results suggested that cPLAϵ is responsible for Ca-dependent formation of NAPEs in the brain as well as the accumulation of NAPEs and NAEs during ischemia, while other enzyme(s) appeared to be involved in the maintenance of basal NAPE levels.","null","null","2022-08-18","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","null","null","159222","159222","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2022.159222","1879-2618","null","null","null","null","null"
"Lysophosphatidic acid, ceramide 1-phosphate and sphingosine 1-phosphate in peripheral blood of patients with idiophathic pulmonary fibrosis","Lysophosphatidic acid, ceramide 1-phosphate and sphingosine 1-phosphate in peripheral blood of patients with idiophathic pulmonary fibrosis","Tamotsu Tanaka, Kazuya Koyama, Naoko Takahashi, Katsuya Morito, Hanif Ali, Momoyo Azuma, Kozo Kagawa, Hiroshi Kawano, Rumana Yesmin, Mutsumi Aihara, Yasuhiko Nishioka","Tamotsu Tanaka, Kazuya Koyama, Naoko Takahashi, Katsuya Morito, Hanif Ali, Momoyo Azuma, Kozo Kagawa, Hiroshi Kawano, Rumana Yesmin, Mutsumi Aihara, Yasuhiko Nishioka","null","Idiopathic pulmonary fibrosis (IPF) is the most common idiopathic interstitial pneumonias. Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) are signaling lipids that evoke growth factor-like responses to many cells. Recent studies revealed the involvement of LPA and S1P in the pathology of IPF. In this study, we determined LPA, S1P and ceramide 1-phosphate (C1P) in peripheral blood plasma of IPF patients, and examined correlation to the vital capacity of lung (VC), an indicator of development of fibrosis. Blood plasma samples were taken from eleven patients with IPF and seven healthy volunteers. The lipids of the sample were extracted and subjected to liquid chromatography-tandem mass spectrometry for analysis. Results showed that there is a significant negative correlation between VC and plasma LPA levels, indicating that IPF patients with advanced fibrosis had higher concentration of LPA in their plasma. Average of S1P levels were significantly higher in IPF patients than those in healthy subjects. Although it is not statistically significant, a similar correlation trend that observed in LPA levels also found between VC and S1P levels. These results indicated that plasma LPA and S1P may be associated with deterioration of pulmonary function of IPF patients. J. Med. Invest. 69 : 196-203, August, 2022.","Idiopathic pulmonary fibrosis (IPF) is the most common idiopathic interstitial pneumonias. Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) are signaling lipids that evoke growth factor-like responses to many cells. Recent studies revealed the involvement of LPA and S1P in the pathology of IPF. In this study, we determined LPA, S1P and ceramide 1-phosphate (C1P) in peripheral blood plasma of IPF patients, and examined correlation to the vital capacity of lung (VC), an indicator of development of fibrosis. Blood plasma samples were taken from eleven patients with IPF and seven healthy volunteers. The lipids of the sample were extracted and subjected to liquid chromatography-tandem mass spectrometry for analysis. Results showed that there is a significant negative correlation between VC and plasma LPA levels, indicating that IPF patients with advanced fibrosis had higher concentration of LPA in their plasma. Average of S1P levels were significantly higher in IPF patients than those in healthy subjects. Although it is not statistically significant, a similar correlation trend that observed in LPA levels also found between VC and S1P levels. These results indicated that plasma LPA and S1P may be associated with deterioration of pulmonary function of IPF patients. J. Med. Invest. 69 : 196-203, August, 2022.","null","null","2022","The Journal of Medical Investigation : JMI","The Journal of Medical Investigation : JMI","Vol.69","No.3.4","196","203","eng","true","null","scientific_journal","null","null","10.2152/jmi.69.196","1349-6867","null","null","null","null","null"
"Characterization of uptake and metabolism of very long-chain fatty acids in peroxisome-deficient CHO cells","Characterization of uptake and metabolism of very long-chain fatty acids in peroxisome-deficient CHO cells","Ali Hanif, Morito Katsuya, Rumana Hasi Yesmin, Mutsumi Aihara, Junji Hayashi, Ryushi Kawakami, Kaori Kanemaru, Koichiro Tsuchiya, Sango Kazunori, Tamotsu Tanaka","Ali Hanif, Morito Katsuya, Rumana Hasi Yesmin, Mutsumi Aihara, Junji Hayashi, Ryushi Kawakami, Kaori Kanemaru, Koichiro Tsuchiya, Sango Kazunori, Tamotsu Tanaka","null","Fatty acids (FAs) longer than C20 are classified as very long-chain fatty acids (VLCFAs). Although biosynthesis and degradation of VLCFAs are important for the development and integrity of the myelin sheath, knowledge on the incorporation of extracellular VLCFAs into the cells is limited due to the experimental difficulty of solubilizing them. In this study, we found that a small amount of isopropanol solubilized VLCFAs in aqueous medium by facilitating the formation of the VLCFA/albumin complex. Using this solubilizing technique, we examined the role of the peroxisome in the uptake and metabolism of VLCFAs in Chinese hamster ovary (CHO) cells. When wild-type CHO cells were incubated with saturated VLCFAs (S-VLCFAs), such as C23:0 FA, C24:0 FA, and C26:0 FA, extensive uptake was observed. Most of the incorporated S-VLCFAs were oxidatively degraded without acylation into cellular lipids. In contrast, in peroxisome-deficient CHO cells uptake of S-VLCFAs was marginal and oxidative metabolism was not observed. Extensive uptake and acylation of monounsaturated (MU)-VLCFAs, such as C24:1 FA and C22:1 FA, were observed in both types of CHO cells. However, oxidative metabolism was evident only in wild-type cells. Similar manners of uptake and metabolism of S-VLCFAs and MU-VLCFAs were observed in IFRS1, a Schwan cell-derived cell line. These results indicate that peroxisome-deficient cells limit intracellular S-VLCFAs at a low level by halting uptake, and as a result, peroxisome-deficient cells almost completely lose the clearance ability of S-VLCFAs accumulated outside of the cells.","Fatty acids (FAs) longer than C20 are classified as very long-chain fatty acids (VLCFAs). Although biosynthesis and degradation of VLCFAs are important for the development and integrity of the myelin sheath, knowledge on the incorporation of extracellular VLCFAs into the cells is limited due to the experimental difficulty of solubilizing them. In this study, we found that a small amount of isopropanol solubilized VLCFAs in aqueous medium by facilitating the formation of the VLCFA/albumin complex. Using this solubilizing technique, we examined the role of the peroxisome in the uptake and metabolism of VLCFAs in Chinese hamster ovary (CHO) cells. When wild-type CHO cells were incubated with saturated VLCFAs (S-VLCFAs), such as C23:0 FA, C24:0 FA, and C26:0 FA, extensive uptake was observed. Most of the incorporated S-VLCFAs were oxidatively degraded without acylation into cellular lipids. In contrast, in peroxisome-deficient CHO cells uptake of S-VLCFAs was marginal and oxidative metabolism was not observed. Extensive uptake and acylation of monounsaturated (MU)-VLCFAs, such as C24:1 FA and C22:1 FA, were observed in both types of CHO cells. However, oxidative metabolism was evident only in wild-type cells. Similar manners of uptake and metabolism of S-VLCFAs and MU-VLCFAs were observed in IFRS1, a Schwan cell-derived cell line. These results indicate that peroxisome-deficient cells limit intracellular S-VLCFAs at a low level by halting uptake, and as a result, peroxisome-deficient cells almost completely lose the clearance ability of S-VLCFAs accumulated outside of the cells.","null","null","2022","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1867","No.2","159088","159088","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2021.159088","1879-2618","null","null","null","null","null"
"Involvement of acid ceramidase in the degradation of bioactive N-acylethanolamines","Involvement of acid ceramidase in the degradation of bioactive N-acylethanolamines","Kazuhito Tsuboi, Tatsuya Tai, Ryouhei Yamashita, Hanif Ali, Takashi Watanabe, Toru Uyama, Yoko Okamoto, Keisuke Kitakaze, Yasuhiro Takenouchi, Shinji Go, Iffat Sonia Ara Rahman, Hitoshi Houchi, Tamotsu Tanaka, Yasuo Okamoto, Akira Tokumura, Junko Matsuda, Natsuo Ueda","Kazuhito Tsuboi, Tatsuya Tai, Ryouhei Yamashita, Hanif Ali, Takashi Watanabe, Toru Uyama, Yoko Okamoto, Keisuke Kitakaze, Yasuhiro Takenouchi, Shinji Go, Iffat Sonia Ara Rahman, Hitoshi Houchi, Tamotsu Tanaka, Yasuo Okamoto, Akira Tokumura, Junko Matsuda, Natsuo Ueda","null","Bioactive N-acylethanolamines (NAEs) include palmitoylethanolamide, oleoylethanolamide, and anandamide, which exert anti-inflammatory, anorexic, and cannabimimetic actions, respectively. The degradation of NAEs has been attributed to two hydrolases, fatty acid amide hydrolase and NAE acid amidase (NAAA). Acid ceramidase (AC) is a lysosomal enzyme that hydrolyzes ceramide (N-acylsphingosine), which resembles NAAA in structure and function. In the present study, we examined the role of AC in the degradation of NAEs. First, we demonstrated that purified recombinant human AC can hydrolyze various NAEs with lauroylethanolamide (C12:0-NAE) as the most reactive NAE substrate. We then used HEK293 cells metabolically labeled with [C]ethanolamine, and revealed that overexpressed AC lowered the levels of C-labeled NAE. As analyzed with liquid chromatography-tandem mass spectrometry, AC overexpression decreased the amounts of different NAE species. Furthermore, suppression of endogenous AC in LNCaP prostate cells by siRNA increased the levels of various NAEs. Lastly, tissue homogenates from mice genetically lacking saposin D, a presumable activator protein of AC, showed much lower hydrolyzing activity for NAE as well as ceramide than the homogenates from wild-type mice. These results demonstrate the ability of AC to hydrolyze NAEs and suggest its physiological role as a third NAE hydrolase.","Bioactive N-acylethanolamines (NAEs) include palmitoylethanolamide, oleoylethanolamide, and anandamide, which exert anti-inflammatory, anorexic, and cannabimimetic actions, respectively. The degradation of NAEs has been attributed to two hydrolases, fatty acid amide hydrolase and NAE acid amidase (NAAA). Acid ceramidase (AC) is a lysosomal enzyme that hydrolyzes ceramide (N-acylsphingosine), which resembles NAAA in structure and function. In the present study, we examined the role of AC in the degradation of NAEs. First, we demonstrated that purified recombinant human AC can hydrolyze various NAEs with lauroylethanolamide (C12:0-NAE) as the most reactive NAE substrate. We then used HEK293 cells metabolically labeled with [C]ethanolamine, and revealed that overexpressed AC lowered the levels of C-labeled NAE. As analyzed with liquid chromatography-tandem mass spectrometry, AC overexpression decreased the amounts of different NAE species. Furthermore, suppression of endogenous AC in LNCaP prostate cells by siRNA increased the levels of various NAEs. Lastly, tissue homogenates from mice genetically lacking saposin D, a presumable activator protein of AC, showed much lower hydrolyzing activity for NAE as well as ceramide than the homogenates from wild-type mice. These results demonstrate the ability of AC to hydrolyze NAEs and suggest its physiological role as a third NAE hydrolase.","null","null","2021-09","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1866","No.9","158972","158972","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2021.158972","1879-2618","null","null","null","null","null"
"Massspectrometric analysis of sphingomyelin with N-alfa-hydroxy fatty acyl residue in mouse tissues","Massspectrometric analysis of sphingomyelin with N-alfa-hydroxy fatty acyl residue in mouse tissues","Hanif Ali, Ryouhei Yamashita, Jun-ichi Morishige, Katsuya Morito, Naoya Kakiuchi, Junji Hayashi, Mutsumi Aihara, Ryushi Kawakami, Koichiro Tsuchiya, Tamotsu Tanaka","Hanif Ali, Ryouhei Yamashita, Jun-ichi Morishige, Katsuya Morito, Naoya Kakiuchi, Junji Hayashi, Mutsumi Aihara, Ryushi Kawakami, Koichiro Tsuchiya, Tamotsu Tanaka","null","null","null","null","null","2021-09","Lipids","Lipids","Vol.56","No.2","181","188","eng","true","null","scientific_journal","null","null","10.1002/lipd.12285","1558-9307","null","http://dx.doi.org/10.1002/lipd.12285","null","null","null"
"Distinct contributions of two choline-producing enzymatic activities to lysophosphatidic acid production in human amniotic fluid from pregnant women in the second trimester and after parturition","Distinct contributions of two choline-producing enzymatic activities to lysophosphatidic acid production in human amniotic fluid from pregnant women in the second trimester and after parturition","Midori Fukui, Toshihiko Tsutsumi, Aimi Yamamoto-Mikami, Katsuya Morito, Naoko Takahashi, Tamotsu Tanaka, Tekeshi Iwasa, Akira Kuwahara, Minoru Irahara, Akira Tokumura","Midori Fukui, Toshihiko Tsutsumi, Aimi Yamamoto-Mikami, Katsuya Morito, Naoko Takahashi, Tamotsu Tanaka, Tekeshi Iwasa, Akira Kuwahara, Minoru Irahara, Akira Tokumura","null","null","null","null","null","2020-10","Prostaglandins & Other Lipid Mediators","Prostaglandins & Other Lipid Mediators","Vol.150","null","106471","106471","eng","true","null","scientific_journal","null","null","10.1016/j.prostaglandins.2020.106471","1098-8823","null","null","null","null","null"
"Identification of human glycerophosphodiesterase 3 as an ectophospholipase C that converts the G protein-coupled receptor 55 agonist lysophosphatidylinositol to bioactive monoacylglycerols in cultured mammalian cells.","Identification of human glycerophosphodiesterase 3 as an ectophospholipase C that converts the G protein-coupled receptor 55 agonist lysophosphatidylinositol to bioactive monoacylglycerols in cultured mammalian cells.","Toshihiko Tsutsumi, Risa Matsuda, Katsuya Morito, Kohei Kawabata, Miho Yokota, Miki Nikawadori, Manami Inoue-Fujiwara, Satoshi Kawashima, Mayumi Hidaka, Takenori Yamamoto, Naoshi Yamazaki, Tamotsu Tanaka, Yasuo Shinohara, Hiroyuki Nishi, Akira Tokumura","Toshihiko Tsutsumi, Risa Matsuda, Katsuya Morito, Kohei Kawabata, Miho Yokota, Miki Nikawadori, Manami Inoue-Fujiwara, Satoshi Kawashima, Mayumi Hidaka, Takenori Yamamoto, Naoshi Yamazaki, Tamotsu Tanaka, Yasuo Shinohara, Hiroyuki Nishi, Akira Tokumura","null","null","null","null","null","2020-09","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1865","No.9","158761","158761","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2020.158761","1388-1981","null","null","null","null","null"
"Isolation of glycosylinositol phosphoceramide and phytoceramide 1-phosphate in plants and their chemical stabilities.","Isolation of glycosylinositol phosphoceramide and phytoceramide 1-phosphate in plants and their chemical stabilities.","Rumana Yesmin Hasi, Dai Majima, Katsuya Morito, Hanif Ali, Kentaro Kogure, Meera Nanjundan, Junji Hayashi, Ryushi Kawakami, Kaori Kanemaru, Tamotsu Tanaka","Rumana Yesmin Hasi, Dai Majima, Katsuya Morito, Hanif Ali, Kentaro Kogure, Meera Nanjundan, Junji Hayashi, Ryushi Kawakami, Kaori Kanemaru, Tamotsu Tanaka","null","Glycosylinositol phosphoceramide (GIPC) is a sphingophospholipid in plants. Recently, we identified that GIPC is hydrolyzed to phytoceramide 1-phosphate (PC1P) by an uncharacterized phospholipase D activity following homogenization of certain plant tissues. We now developed methods for isolation of GIPC and PC1P from plant tissues and characterized their chemical stabilities. Hydrophilic solvents, namely a lower layer of a mixed solvent system consisting of isopropanol/hexane/water (55:20:25, v/v/v) was efficient solvent for extraction and eluent in column chromatography. GIPC was isolated by Sephadex column chromatography followed by TLC. A conventional method, such as the Bligh and Dyer method, was applicable for PC1P extraction. Specifically, PC1P was isolated by TLC following mild alkali treatment of lipid extracts of plants. The yields of GIPC and PC1P in our methods were both around 50-70%. We found that PC1P is tolerant against heat (up to 125 °C), strong acid (up to 10 M HCl), and mild alkali (0.1 M KOH). In contrast, significant degradation of GIPC occurred at 100 °C and 1.0 M HCl treatment, suggesting the instability of the inositol glycan moiety in these conditions. These data will be useful for further biochemical and nutritional studies on these sphingolipids.","Glycosylinositol phosphoceramide (GIPC) is a sphingophospholipid in plants. Recently, we identified that GIPC is hydrolyzed to phytoceramide 1-phosphate (PC1P) by an uncharacterized phospholipase D activity following homogenization of certain plant tissues. We now developed methods for isolation of GIPC and PC1P from plant tissues and characterized their chemical stabilities. Hydrophilic solvents, namely a lower layer of a mixed solvent system consisting of isopropanol/hexane/water (55:20:25, v/v/v) was efficient solvent for extraction and eluent in column chromatography. GIPC was isolated by Sephadex column chromatography followed by TLC. A conventional method, such as the Bligh and Dyer method, was applicable for PC1P extraction. Specifically, PC1P was isolated by TLC following mild alkali treatment of lipid extracts of plants. The yields of GIPC and PC1P in our methods were both around 50-70%. We found that PC1P is tolerant against heat (up to 125 °C), strong acid (up to 10 M HCl), and mild alkali (0.1 M KOH). In contrast, significant degradation of GIPC occurred at 100 °C and 1.0 M HCl treatment, suggesting the instability of the inositol glycan moiety in these conditions. These data will be useful for further biochemical and nutritional studies on these sphingolipids.","null","null","2020-06-12","Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences","Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences","Vol.1152","null","122213","122213","eng","true","null","scientific_journal","null","null","10.1016/j.jchromb.2020.122213","1873-376X","null","null","null","null","null"
"A low level of lysophosphatidic acid in human gingival crevicular fluid from patients with periodontitis due to high soluble lysophospholipase activity: Its potential protective role on alveolar bone loss by periodontitis.","A low level of lysophosphatidic acid in human gingival crevicular fluid from patients with periodontitis due to high soluble lysophospholipase activity: Its potential protective role on alveolar bone loss by periodontitis.","Tohru Hashimura, Jun-ichi Kido, Risa Matsuda, Miho Yokota, Hirokazu Matsui, Manami Inoue-Fujiwara, Yuji Inagaki, Mayumi Hidaka, Tamotsu Tanaka, Toshihiko Tsutsumi, Toshihiko Nagata, Akira Tokumura","Tohru Hashimura, Jun-ichi Kido, Risa Matsuda, Miho Yokota, Hirokazu Matsui, Manami Inoue-Fujiwara, Yuji Inagaki, Mayumi Hidaka, Tamotsu Tanaka, Toshihiko Tsutsumi, Toshihiko Nagata, Akira Tokumura","null","We previously detected a submicromolar concentration of lysophosphatidic acid (LPA) in human saliva. Here, we compare LPA concentrations in human gingival crevicular fluid (GCF) from patients with periodontitis and healthy controls, and examine how the local LPA levels are regulated enzymatically. The concentrations of LPA and its precursor lysophospholipids in GCF was measured by liquid chromatography-tandem mass spectrometry. The LPA-producing and LPA-degrading enzymatic activities were measured by quantifying the liberated choline and free fatty acid, respectively. The concentration of LPA in GCF of periodontitis patients was lower than that of healthy controls, due to higher soluble lysophospholipase activity toward LPA. LPA was found to prevent survival of Sa3, a human gingival epithelium-derived tumor cell line, activate Sa3 through Ca mobilization, and release interleukin 6 from Sa3 in vitro. Furthermore, local injection of LPA into the gingiva attenuated ligature-induced experimental alveolar bone loss induced by oral bacteria inoculation in a rat model of periodontitis in vivo. A high concentration of LPA in human GCF is necessary to maintain normal gingival epithelial integrity and function, protecting the progression of periodontitis.","We previously detected a submicromolar concentration of lysophosphatidic acid (LPA) in human saliva. Here, we compare LPA concentrations in human gingival crevicular fluid (GCF) from patients with periodontitis and healthy controls, and examine how the local LPA levels are regulated enzymatically. The concentrations of LPA and its precursor lysophospholipids in GCF was measured by liquid chromatography-tandem mass spectrometry. The LPA-producing and LPA-degrading enzymatic activities were measured by quantifying the liberated choline and free fatty acid, respectively. The concentration of LPA in GCF of periodontitis patients was lower than that of healthy controls, due to higher soluble lysophospholipase activity toward LPA. LPA was found to prevent survival of Sa3, a human gingival epithelium-derived tumor cell line, activate Sa3 through Ca mobilization, and release interleukin 6 from Sa3 in vitro. Furthermore, local injection of LPA into the gingiva attenuated ligature-induced experimental alveolar bone loss induced by oral bacteria inoculation in a rat model of periodontitis in vivo. A high concentration of LPA in human GCF is necessary to maintain normal gingival epithelial integrity and function, protecting the progression of periodontitis.","null","null","2020-03-13","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1865","No.7","158698","158698","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2020.158698","1388-1981","null","null","null","null","null"
"Characteristics of unique endocytosis induced by weak current for cytoplasmic drug delivery","Characteristics of unique endocytosis induced by weak current for cytoplasmic drug delivery","Tasuku Torao, Miyuki Mimura, Yasufumi Ohshima, Kohki Fujikawa, Mahadi Hasan, Tatsuharu Shimokawa, Naoshi Yamazaki, Hidenori ANDO, Tatsuhiro Ishida, Tatsuya Fukuta, Tamotsu Tanaka, Kentaro Kogure","Tasuku Torao, Miyuki Mimura, Yasufumi Ohshima, Kohki Fujikawa, Mahadi Hasan, Tatsuharu Shimokawa, Naoshi Yamazaki, Hidenori ANDO, Tatsuhiro Ishida, Tatsuya Fukuta, Tamotsu Tanaka, Kentaro Kogure","null","null","null","null","null","2020-02","International Journal of Pharmaceutics","International Journal of Pharmaceutics","Vol.576","null","119010","119010","eng","true","null","scientific_journal","null","null","10.1016/j.ijpharm.2019.119010","0378-5173","null","null","null","null","null"
"Intracellular Ca2+-dependent formation of N-acyl-phosphatidylethanolamines by human cytosolic phospholipase A2ϵ.","Intracellular Ca2+-dependent formation of N-acyl-phosphatidylethanolamines by human cytosolic phospholipase A2ϵ.","Smriti Binte Sultana Mustafiz, Toru Uyama, Katsuya Morito, Naoko Takahashi, Katsuhisa Kawai, Zahir Hussain, Kazuhito Tsuboi, Nobukazu Araki, Kei Yamamoto, Tamotsu Tanaka, Natsuo Ueda","Smriti Binte Sultana Mustafiz, Toru Uyama, Katsuya Morito, Naoko Takahashi, Katsuhisa Kawai, Zahir Hussain, Kazuhito Tsuboi, Nobukazu Araki, Kei Yamamoto, Tamotsu Tanaka, Natsuo Ueda","null","null","null","null","null","2019-12","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1864","No.12","158515","158515","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2019.158515","1388-1981","null","null","null","null","null"
"Gut microbial metabolites of linoleic acid are metabolized by accelerated peroxisomal β-oxidation in mammalian cells","Gut microbial metabolites of linoleic acid are metabolized by accelerated peroxisomal β-oxidation in mammalian cells","Katsuya Morito, Ryota Shimizu, Nahoko Kitamura, Si-Bum Park, Shigenobu Kishino, Jun Ogawa, Tatsuya Fukuta, Kentaro Kogure, Tamotsu Tanaka","Katsuya Morito, Ryota Shimizu, Nahoko Kitamura, Si-Bum Park, Shigenobu Kishino, Jun Ogawa, Tatsuya Fukuta, Kentaro Kogure, Tamotsu Tanaka","null","Microorganisms in animal gut produce unusual fatty acids from the ingested diet. Two types of hydroxy fatty acids (HFAs), 10-hydroxy-cis-12-octadecenoic acid (HYA) and 10-hydroxy-octadecanoic acid (HYB), are linoleic acid (LA) metabolites produced by Lactobacillus plantarum. In this study, we investigated the metabolism of these HFAs in mammalian cells. When Chinese hamster ovary (CHO) cells were cultured with HYA, approximately 50% of the supplemented HYA disappeared from the dish within 24 h. On the other hand, the amount of HYA that disappeared from the dish of peroxisome (PEX)-deficient CHO cells was lower than 20%. Significant amounts of C2- and C4-chain-shortened metabolites of HYA were detected in culture medium of HYA-supplemented CHO cells, but not in medium of PEX-deficient cells. These results suggested that peroxisomal β-oxidation is involved in the disappearance of HYA. The PEX-dependent disappearance was observed in the experiment with HYB, but not with LA. We also found that HYA treatment up-regulates peroxisomal β-oxidation activity of human gastric MKN74 cells and intestinal Caco-2 cells. These results indicate a possibility that HFAs produced from gut bacteria affect lipid metabolism of host via modulation of peroxisomal β-oxidation activity.","Microorganisms in animal gut produce unusual fatty acids from the ingested diet. Two types of hydroxy fatty acids (HFAs), 10-hydroxy-cis-12-octadecenoic acid (HYA) and 10-hydroxy-octadecanoic acid (HYB), are linoleic acid (LA) metabolites produced by Lactobacillus plantarum. In this study, we investigated the metabolism of these HFAs in mammalian cells. When Chinese hamster ovary (CHO) cells were cultured with HYA, approximately 50% of the supplemented HYA disappeared from the dish within 24 h. On the other hand, the amount of HYA that disappeared from the dish of peroxisome (PEX)-deficient CHO cells was lower than 20%. Significant amounts of C2- and C4-chain-shortened metabolites of HYA were detected in culture medium of HYA-supplemented CHO cells, but not in medium of PEX-deficient cells. These results suggested that peroxisomal β-oxidation is involved in the disappearance of HYA. The PEX-dependent disappearance was observed in the experiment with HYB, but not with LA. We also found that HYA treatment up-regulates peroxisomal β-oxidation activity of human gastric MKN74 cells and intestinal Caco-2 cells. These results indicate a possibility that HFAs produced from gut bacteria affect lipid metabolism of host via modulation of peroxisomal β-oxidation activity.","null","null","2019-11","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1864","No.11","1619","1628","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2019.07.010","1879-2618","null","null","null","null","null"
"Quantitative Analysis of Glycosylinositol Phosphoceramide and Phytoceramide 1-Phosphate in Vegetables","Quantitative Analysis of Glycosylinositol Phosphoceramide and Phytoceramide 1-Phosphate in Vegetables","Rumana Yesmin Hasi, Makoto Miyagi, Takashi Kida, Tatsuya Fukuta, Kentaro Kogure, Junji Hayashi, Ryushi Kawakami, Kaori Kanemaru, Tamotsu Tanaka","Rumana Yesmin Hasi, Makoto Miyagi, Takashi Kida, Tatsuya Fukuta, Kentaro Kogure, Junji Hayashi, Ryushi Kawakami, Kaori Kanemaru, Tamotsu Tanaka","null","Previously, we found an unidentified sphingolipid in cabbage, and determined it as phytoceramide 1-phosphate (PC1P). PC1P is found to be produced from glycosylinositol phosphoceramide (GIPC) by the action of phospholipase D (PLD) activity. Although GIPC is abundant sphingolipid, especially in cruciferous vegetables, amount of daily intake, digestibility and nutritional activity of GIPC are not well understood. Here, we investigated amounts of GIPC and PC1P in vegetables. GIPC was found in all vegetables examined (13 kinds) at levels 3-20 mg/100 g (wet weight). On the other hand, PC1P was present in limited vegetables which show higher GIPC-PLD activity, such as inner cabbage leaves (5.2 mg/100 g). Because PC1P is formed during homogenization by activated GIPC-PLD, level of PC1P in boiled cabbage leaves was very low. Although digestibility of GIPC is unknown at present, a portion of dietary GIPC is considered to be converted to PC1P during mastication by plant-derived GIPC-PLD activity in some vegetables.","Previously, we found an unidentified sphingolipid in cabbage, and determined it as phytoceramide 1-phosphate (PC1P). PC1P is found to be produced from glycosylinositol phosphoceramide (GIPC) by the action of phospholipase D (PLD) activity. Although GIPC is abundant sphingolipid, especially in cruciferous vegetables, amount of daily intake, digestibility and nutritional activity of GIPC are not well understood. Here, we investigated amounts of GIPC and PC1P in vegetables. GIPC was found in all vegetables examined (13 kinds) at levels 3-20 mg/100 g (wet weight). On the other hand, PC1P was present in limited vegetables which show higher GIPC-PLD activity, such as inner cabbage leaves (5.2 mg/100 g). Because PC1P is formed during homogenization by activated GIPC-PLD, level of PC1P in boiled cabbage leaves was very low. Although digestibility of GIPC is unknown at present, a portion of dietary GIPC is considered to be converted to PC1P during mastication by plant-derived GIPC-PLD activity in some vegetables.","null","null","2019-10-11","Journal of Nutritional Science and Vitaminology","Journal of Nutritional Science and Vitaminology","Vol.65","No.Supplement","S175","S179","eng","true","null","scientific_journal","null","null","10.3177/jnsv.65.S175","1881-7742","null","null","null","null","null"
"Glycosylinositol phosphoceramide-specific phospholipase D activity catalyzes transphosphatidylation","Glycosylinositol phosphoceramide-specific phospholipase D activity catalyzes transphosphatidylation","Hasi Yesmin Rumana, Makoto Miyagi, Katsuya Morito, Toshiki Ishikawa, Maki Kawai-Yamada, Hiroyuki Imai, Tatsuya Fukuta, Kentaro Kogure, Kaori Kanemaru, Junji Hayashi, Ryushi Kawakami, Tamotsu Tanaka","Hasi Yesmin Rumana, Makoto Miyagi, Katsuya Morito, Toshiki Ishikawa, Maki Kawai-Yamada, Hiroyuki Imai, Tatsuya Fukuta, Kentaro Kogure, Kaori Kanemaru, Junji Hayashi, Ryushi Kawakami, Tamotsu Tanaka","null","Glycosylinositol phosphoceramide (GIPC) is the most abundant sphingolipid in plants and fungi. Recently, we detected GIPC-specific phospholipase D (GIPC-PLD) activity in plants. Here, we found that GIPC-PLD activity in young cabbage leaves catalyzes transphosphatidylation. The available alcohol for this reaction is a primary alcohol with a chain length below C4. Neither secondary alcohol, tertiary alcohol, choline, serine nor glycerol serves as an acceptor for transphosphatidylation of GIPC-PLD. We also found that cabbage GIPC-PLD prefers GIPC containing two sugars. Neither inositol phosphoceramide, mannosylinositol phosphoceramide nor GIPC with three sugar chains served as substrate. GIPC-PLD will become a useful catalyst for modification of polar head group of sphingophospholipid.","Glycosylinositol phosphoceramide (GIPC) is the most abundant sphingolipid in plants and fungi. Recently, we detected GIPC-specific phospholipase D (GIPC-PLD) activity in plants. Here, we found that GIPC-PLD activity in young cabbage leaves catalyzes transphosphatidylation. The available alcohol for this reaction is a primary alcohol with a chain length below C4. Neither secondary alcohol, tertiary alcohol, choline, serine nor glycerol serves as an acceptor for transphosphatidylation of GIPC-PLD. We also found that cabbage GIPC-PLD prefers GIPC containing two sugars. Neither inositol phosphoceramide, mannosylinositol phosphoceramide nor GIPC with three sugar chains served as substrate. GIPC-PLD will become a useful catalyst for modification of polar head group of sphingophospholipid.","null","null","2019-08-26","The Journal of Biochemistry","The Journal of Biochemistry","Vol.166","No.5","441","448","eng","true","null","scientific_journal","null","null","10.1093/jb/mvz056","1756-2651","null","null","null","null","null"
"Leukocyte-mimetic liposomes possessing leukocyte membrane proteins pass through inflamed endothelial cell layer by regulating intercellular junctions","Leukocyte-mimetic liposomes possessing leukocyte membrane proteins pass through inflamed endothelial cell layer by regulating intercellular junctions","Tatsuya Fukuta, Shintaroh Yoshimi, Tamotsu Tanaka, Kentaro Kogure","Tatsuya Fukuta, Shintaroh Yoshimi, Tamotsu Tanaka, Kentaro Kogure","null","Nanoparticles such as liposomes have been applied for the treatment of various diseases such as cancer and inflammatory diseases by utilizing the enhanced permeability and retention effect. However, their entry into inflammation sites is still limited since passive delivery of nanoparticles is often hampered by the presence of endothelial barriers. As leukocytes can pass through the inflamed endothelium via utilizing membrane protein functions, we hypothesized that incorporating leukocyte membrane proteins onto liposomal membranes may impart leukocyte-mimicking functions to liposomes, allowing for their adherence to and active passage through the inflamed endothelium. Herein, we developed leukocyte-mimetic liposomes (LM-Lipo) by leukocyte membrane protein transfer and evaluated their function in vitro. Transfer of membrane proteins from human leukemia cells onto liposomal membranes allowed for significant association of the liposomes with inflamed human endothelial cells, and subsequent passage through inflamed endothelial cell layer. The confocal images showed that LM-Lipo significantly induced vascular endothelial-cadherin displacement. These results indicate that LM-Lipo adhered to and regulated intercellular junctions of inflamed endothelial cell layer, resulting in passage through the layer, by mimicking the function of leukocytes. Furthermore, it is suggested that liposomes possessing leukocyte-like functions could be useful for drug delivery to inflammation sites by overcoming endothelial barriers.","Nanoparticles such as liposomes have been applied for the treatment of various diseases such as cancer and inflammatory diseases by utilizing the enhanced permeability and retention effect. However, their entry into inflammation sites is still limited since passive delivery of nanoparticles is often hampered by the presence of endothelial barriers. As leukocytes can pass through the inflamed endothelium via utilizing membrane protein functions, we hypothesized that incorporating leukocyte membrane proteins onto liposomal membranes may impart leukocyte-mimicking functions to liposomes, allowing for their adherence to and active passage through the inflamed endothelium. Herein, we developed leukocyte-mimetic liposomes (LM-Lipo) by leukocyte membrane protein transfer and evaluated their function in vitro. Transfer of membrane proteins from human leukemia cells onto liposomal membranes allowed for significant association of the liposomes with inflamed human endothelial cells, and subsequent passage through inflamed endothelial cell layer. The confocal images showed that LM-Lipo significantly induced vascular endothelial-cadherin displacement. These results indicate that LM-Lipo adhered to and regulated intercellular junctions of inflamed endothelial cell layer, resulting in passage through the layer, by mimicking the function of leukocytes. Furthermore, it is suggested that liposomes possessing leukocyte-like functions could be useful for drug delivery to inflammation sites by overcoming endothelial barriers.","null","null","2019-05","International Journal of Pharmaceutics","International Journal of Pharmaceutics","Vol.563","null","314","323","eng","true","null","scientific_journal","null","null","10.1016/j.ijpharm.2019.04.027","1873-3476","null","null","null","null","null"
"Efficacy of high-affinity liposomal astaxanthin on up-regulation of age-related markers induced by oxidative stress in human corneal epithelial cells","Efficacy of high-affinity liposomal astaxanthin on up-regulation of age-related markers induced by oxidative stress in human corneal epithelial cells","Tatsuharu Shimokawa, Mai Yoshida, Tatsuya Fukuta, Tamotsu Tanaka, Toshio Inagi, Kentaro Kogure","Tatsuharu Shimokawa, Mai Yoshida, Tatsuya Fukuta, Tamotsu Tanaka, Toshio Inagi, Kentaro Kogure","null","Decreases in tear volume, unstable tear films and excessive tear evaporation are known to cause desiccation and hyperosmolar stress. These, in turn, induce oxidative stress that is thought to cause dry eye, which is also considered to be age-related disease. We hypothesized that oxidative stress induces up-regulation of age-related markers, and that the antioxidant astaxanthin prepared as a liposomal formulation may be a candidate for the treatment of dry eye. Herein, we examined age-related markers in an dry eye model, and evaluated the efficacy of high-affinity liposomes containing astaxanthin. The dry eye model showed desiccation time-dependent increases in reactive oxygen species. We confirmed the up-regulation of p53, p21 and p16 as a function of desiccation time. Pretreatment with both neutral and slightly-positively-charged astaxanthin liposomal formulations showed significant suppression of up-regulation of all markers, with the positively-charged liposomes exhibiting the greatest efficacy. Furthermore, positively-charged liposomes labeled with fluorescent dyes demonstrated much higher affinity to normal human corneal epithelial cells (HCECs) than neutral liposomes. Taken together, we confirmed the up-regulation of age-related markers, especially p16, in an dry eye model, and demonstrated the potential of high-affinity liposomal astaxanthin for the treatment of dry eye.","Decreases in tear volume, unstable tear films and excessive tear evaporation are known to cause desiccation and hyperosmolar stress. These, in turn, induce oxidative stress that is thought to cause dry eye, which is also considered to be age-related disease. We hypothesized that oxidative stress induces up-regulation of age-related markers, and that the antioxidant astaxanthin prepared as a liposomal formulation may be a candidate for the treatment of dry eye. Herein, we examined age-related markers in an dry eye model, and evaluated the efficacy of high-affinity liposomes containing astaxanthin. The dry eye model showed desiccation time-dependent increases in reactive oxygen species. We confirmed the up-regulation of p53, p21 and p16 as a function of desiccation time. Pretreatment with both neutral and slightly-positively-charged astaxanthin liposomal formulations showed significant suppression of up-regulation of all markers, with the positively-charged liposomes exhibiting the greatest efficacy. Furthermore, positively-charged liposomes labeled with fluorescent dyes demonstrated much higher affinity to normal human corneal epithelial cells (HCECs) than neutral liposomes. Taken together, we confirmed the up-regulation of age-related markers, especially p16, in an dry eye model, and demonstrated the potential of high-affinity liposomal astaxanthin for the treatment of dry eye.","null","null","2019-01","Journal of Clinical Biochemistry and Nutrition","Journal of Clinical Biochemistry and Nutrition","Vol.64","No.1","27","35","eng","true","null","scientific_journal","null","null","10.3164/jcbn.18-27","0912-0009","null","null","null","null","null"
"Addition of high load of lysophosphatidic acid to standard and high-fat chows causes no significant changes of its circulating and peripheral tissue levels but affects body weight and visceral fat mass of mice.","Addition of high load of lysophosphatidic acid to standard and high-fat chows causes no significant changes of its circulating and peripheral tissue levels but affects body weight and visceral fat mass of mice.","Manami Inoue, Yoko Okamoto, Yuta Atsumi, Masatoshi Shiojiri, Mayumi Hidaka, Tamotsu Tanaka, Toshihiko Tsutsumi, Naoki Shirasaka, Akira Tokumura","Manami Inoue, Yoko Okamoto, Yuta Atsumi, Masatoshi Shiojiri, Mayumi Hidaka, Tamotsu Tanaka, Toshihiko Tsutsumi, Naoki Shirasaka, Akira Tokumura","null","Oral administration of lysophosphatidic acid (LPA), a critical intercellular lipid mediator, exerts wound healing and antiulcer effects on gastrointestinal system. To evaluate effects of food-derived LPA on body homeostasis, we measured LPA levels by liquid chromatography-tandem mass spectrometry in chows, feces, plasma, liver, and visceral fat of mice fed a normal or high-fat chow supplemented with or without LPA-rich soybean phospholipids for 30 days. Reductions in daily body weight gains and visceral fat mass were mainly related to lower chow intake by mice fed the LPA-rich high-fat chow, whereas reduced body weight gains and fat mass were mainly related to decreased intestinal triacylglycerol absorption in mice fed LPA-rich chow. Our results showed no significant increase in plasma, liver, or adipose LPA levels, even if a quite high LPA concentration (2.0%) in chows was ingested daily, suggesting limited effects of food-derived LPA on the lumen side of the digestive tract. © 2018 BioFactors, 44(6):548-557, 2018.","Oral administration of lysophosphatidic acid (LPA), a critical intercellular lipid mediator, exerts wound healing and antiulcer effects on gastrointestinal system. To evaluate effects of food-derived LPA on body homeostasis, we measured LPA levels by liquid chromatography-tandem mass spectrometry in chows, feces, plasma, liver, and visceral fat of mice fed a normal or high-fat chow supplemented with or without LPA-rich soybean phospholipids for 30 days. Reductions in daily body weight gains and visceral fat mass were mainly related to lower chow intake by mice fed the LPA-rich high-fat chow, whereas reduced body weight gains and fat mass were mainly related to decreased intestinal triacylglycerol absorption in mice fed LPA-rich chow. Our results showed no significant increase in plasma, liver, or adipose LPA levels, even if a quite high LPA concentration (2.0%) in chows was ingested daily, suggesting limited effects of food-derived LPA on the lumen side of the digestive tract. © 2018 BioFactors, 44(6):548-557, 2018.","null","null","2018-10-16","BioFactors","BioFactors","Vol.44","No.6","548","557","eng","true","null","scientific_journal","null","null","10.1002/biof.1451","1872-8081","null","null","null","null","null"
"Carotenoid Stereochemistry Affects Antioxidative Activity of Liposomes Co-encapsulating Astaxanthin and Tocotrienol","Carotenoid Stereochemistry Affects Antioxidative Activity of Liposomes Co-encapsulating Astaxanthin and Tocotrienol","Misuzu Ishikawa, Shota Hirai, Tatsusada Yoshida, Natsumi Shibuya, Susumu Hama, Yu Takahashi, Tatsuya Fukuta, Tamotsu Tanaka, Shinzo Hosoi, Kentaro Kogure","Misuzu Ishikawa, Shota Hirai, Tatsusada Yoshida, Natsumi Shibuya, Susumu Hama, Yu Takahashi, Tatsuya Fukuta, Tamotsu Tanaka, Shinzo Hosoi, Kentaro Kogure","null","We previously found that antioxidative activity of liposomes co-encapsulating astaxanthin (Asx) and tocotrienols (T3s) was higher than the calculated additive activity, which results from intermolecular interactions between both antioxidants (J. Clin. Biochem. Nutr., 59, 2016, Kamezaki et al.). Herein, we conducted experiments to optimize Asx/α-T3 ratio for high antioxidative activity, and tried to elucidate details of intermolecular interaction of Asx with α-T3. Higher activity than calculated additive value was clearly observed at an Asx/α-T3 ratio of 2 : 1, despite two α-T3 would potentially interact with two terminal rings of one Asx. The synthetic Asx used in this study was a mixture of three stereoisomers, 3R,3'R-form (Asx-R), 3S,3'S-form (Asx-S) and 3R,3'S-meso form (Asx-meso). The calculated binding energy of the Asx-S/α-T3 complex was higher than those of Asx-R/α-T3 and Asx-meso/α-T3, suggesting that Asx-S and α-T3 is the most preferable combination for the intermolecular interaction. The optimal Asx-S/α-T3 ratio for antioxidation was shown to be 1 : 2. These results suggest that the Asx stereochemistry affects the intermolecular interaction of Asx/α-T3. Moreover, the absorption spectrum changes of Asx-S upon co-encapsulation with α-T3 in liposomes indicate that the electronic state of Asx-S is affected by intermolecular interactions with α-T3. Further, intermolecular interactions with α-T3 affected the electronic charges on the C9, C10 and C15 atoms in the polyene moiety of Asx-S. In conclusion, the intermolecular interaction of Asx/T3 depends on the Asx stereochemistry, and caused a change in the electronic state of the Asx polyene moiety by the presence of double bond in the T3 triene moiety.","We previously found that antioxidative activity of liposomes co-encapsulating astaxanthin (Asx) and tocotrienols (T3s) was higher than the calculated additive activity, which results from intermolecular interactions between both antioxidants (J. Clin. Biochem. Nutr., 59, 2016, Kamezaki et al.). Herein, we conducted experiments to optimize Asx/α-T3 ratio for high antioxidative activity, and tried to elucidate details of intermolecular interaction of Asx with α-T3. Higher activity than calculated additive value was clearly observed at an Asx/α-T3 ratio of 2 : 1, despite two α-T3 would potentially interact with two terminal rings of one Asx. The synthetic Asx used in this study was a mixture of three stereoisomers, 3R,3'R-form (Asx-R), 3S,3'S-form (Asx-S) and 3R,3'S-meso form (Asx-meso). The calculated binding energy of the Asx-S/α-T3 complex was higher than those of Asx-R/α-T3 and Asx-meso/α-T3, suggesting that Asx-S and α-T3 is the most preferable combination for the intermolecular interaction. The optimal Asx-S/α-T3 ratio for antioxidation was shown to be 1 : 2. These results suggest that the Asx stereochemistry affects the intermolecular interaction of Asx/α-T3. Moreover, the absorption spectrum changes of Asx-S upon co-encapsulation with α-T3 in liposomes indicate that the electronic state of Asx-S is affected by intermolecular interactions with α-T3. Further, intermolecular interactions with α-T3 affected the electronic charges on the C9, C10 and C15 atoms in the polyene moiety of Asx-S. In conclusion, the intermolecular interaction of Asx/T3 depends on the Asx stereochemistry, and caused a change in the electronic state of the Asx polyene moiety by the presence of double bond in the T3 triene moiety.","null","null","2018-05","Chemical & Pharmaceutical Bulletin","Chemical & Pharmaceutical Bulletin","Vol.66","No.7","714","720","eng","true","null","scientific_journal","null","null","10.1248/cpb.c18-00035","1347-5223","null","null","null","null","null"
"Lysophosphatidic acid in medicinal herbs enhances prostaglandin E2 and protects against indomethacin-induced gastric cell damage in vivo and in vitro","Lysophosphatidic acid in medicinal herbs enhances prostaglandin E2 and protects against indomethacin-induced gastric cell damage in vivo and in vitro","Sheuli Afroz, Ayano Yagi, Kouki Fujikawa, M. Motiur Rahman, Katsuya Morito, Tatsuya Fukuta, Shiro Watanabe, Kazunori Toida, Emi Kiyokage, Taro Shimizu, Tatsuhiro Ishida, Kentaro Kogure, Akira Tokumura, Tamotsu Tanaka","Sheuli Afroz, Ayano Yagi, Kouki Fujikawa, M. Motiur Rahman, Katsuya Morito, Tatsuya Fukuta, Shiro Watanabe, Kazunori Toida, Emi Kiyokage, Taro Shimizu, Tatsuhiro Ishida, Kentaro Kogure, Akira Tokumura, Tamotsu Tanaka","null","Lysophosphatidic acid (LPA) is a bioactive phospholipid that induces diverse biological responses. Recently, we found that LPA ameliorates NSAIDs-induced gastric ulcer in mice. Here, we quantified LPA in 21 medicinal herbs used for treatment of gastrointestinal (GI) disorders. We found that half of them contained LPA at relatively high levels (40-240 μg/g) compared to soybean seed powder (4.6 μg/g), which we previously identified as an LPA-rich food. The LPA in peony (Paeonia lactiflora) root powder is highly concentrated in the lipid fraction that ameliorates indomethacin-induced gastric ulcer in mice. Synthetic 18:1 LPA, peony root LPA and peony root lipid enhanced prostaglandin E production in a gastric cancer cell line, MKN74 cells that express LPA abundantly. These materials also prevented indomethacin-induced cell death and stimulated the proliferation of MKN74 cells. We found that LPA was present in stomach fluids at 2.4 μM, which is an effective LPA concentration for inducing a cellular response in vitro. These results indicated that LPA is one of the active components of medicinal herbs for the treatment of GI disorder and that orally administered LPA-rich herbs may augment the protective actions of endogenous LPA on gastric mucosa.","Lysophosphatidic acid (LPA) is a bioactive phospholipid that induces diverse biological responses. Recently, we found that LPA ameliorates NSAIDs-induced gastric ulcer in mice. Here, we quantified LPA in 21 medicinal herbs used for treatment of gastrointestinal (GI) disorders. We found that half of them contained LPA at relatively high levels (40-240 μg/g) compared to soybean seed powder (4.6 μg/g), which we previously identified as an LPA-rich food. The LPA in peony (Paeonia lactiflora) root powder is highly concentrated in the lipid fraction that ameliorates indomethacin-induced gastric ulcer in mice. Synthetic 18:1 LPA, peony root LPA and peony root lipid enhanced prostaglandin E production in a gastric cancer cell line, MKN74 cells that express LPA abundantly. These materials also prevented indomethacin-induced cell death and stimulated the proliferation of MKN74 cells. We found that LPA was present in stomach fluids at 2.4 μM, which is an effective LPA concentration for inducing a cellular response in vitro. These results indicated that LPA is one of the active components of medicinal herbs for the treatment of GI disorder and that orally administered LPA-rich herbs may augment the protective actions of endogenous LPA on gastric mucosa.","null","null","2018-01-25","Prostaglandins & Other Lipid Mediators","Prostaglandins & Other Lipid Mediators","Vol.135","null","36","44","eng","true","null","scientific_journal","null","null","10.1016/j.prostaglandins.2018.01.003","1098-8823","null","null","null","null","null"
"Peripheral tissue levels and molecular species compositions of N-acyl-phosphatidylethanolamine and its metabolites in mice lacking N-acyl-phosphatidylethanolamine-specific phospholipase D.","Peripheral tissue levels and molecular species compositions of N-acyl-phosphatidylethanolamine and its metabolites in mice lacking N-acyl-phosphatidylethanolamine-specific phospholipase D.","Manami Inoue, Kazuhito Tsuboi, Yoko Okamoto, Mayumi Hidaka, Toru Uyama, Toshihiko Tsutsumi, Tamotsu Tanaka, Natsuo Ueda, Akira Tokumura","Manami Inoue, Kazuhito Tsuboi, Yoko Okamoto, Mayumi Hidaka, Toru Uyama, Toshihiko Tsutsumi, Tamotsu Tanaka, Natsuo Ueda, Akira Tokumura","null","N-acylethanolamines (NAEs), a class of lipid mediators, are produced from N-acyl-phosphatidylethanolamine (NAPE) by several pathways, including the direct release by NAPE-specific phospholipase D (NAPE-PLD) or the multistep pathway via sn-glycero-3-phospho-N-acylethanolamine (Gp-NAE). Using liquid chromatography-tandem mass spectrometry, we compared peripheral tissue levels of NAPE, Gp-NAE and NAE in NAPE-PLD-deficient (NAPE-PLD-/-) and wild type (WT) mice. NAPE-PLD was suggested to play a major role in the NAPE degradation in heart, kidney, and liver, but not in jejunum, because the NAPE levels except jejunum were significantly higher in NAPE-PLD-/- mice than in WT mice. The deletion of NAPE-PLD failed to alter the NAE levels of these tissues, suggesting its limited role in the NAE production. The enzyme assays with tissue homogenates confirmed the presence of NAPE-PLD-independent pathways in these peripheral tissues. Gp-NAE species having an acyl moiety with 22 carbons and 6 double bonds was enriched in these peripheral tissues. As for sn-2 acyl species of NAPE, 18:2-acyl-containing NAPE species were predominant over 18:1-containing species in heart, liver, and jejunum. Our results show that both molecular species composition of NAPE, NAE and Gp-NAE and their dependencies on Napepld are different among the peripheral tissues, suggesting that each tissue has distinct metabolic pathways and these NAE-containing lipids play tissue-specific roles.","N-acylethanolamines (NAEs), a class of lipid mediators, are produced from N-acyl-phosphatidylethanolamine (NAPE) by several pathways, including the direct release by NAPE-specific phospholipase D (NAPE-PLD) or the multistep pathway via sn-glycero-3-phospho-N-acylethanolamine (Gp-NAE). Using liquid chromatography-tandem mass spectrometry, we compared peripheral tissue levels of NAPE, Gp-NAE and NAE in NAPE-PLD-deficient (NAPE-PLD-/-) and wild type (WT) mice. NAPE-PLD was suggested to play a major role in the NAPE degradation in heart, kidney, and liver, but not in jejunum, because the NAPE levels except jejunum were significantly higher in NAPE-PLD-/- mice than in WT mice. The deletion of NAPE-PLD failed to alter the NAE levels of these tissues, suggesting its limited role in the NAE production. The enzyme assays with tissue homogenates confirmed the presence of NAPE-PLD-independent pathways in these peripheral tissues. Gp-NAE species having an acyl moiety with 22 carbons and 6 double bonds was enriched in these peripheral tissues. As for sn-2 acyl species of NAPE, 18:2-acyl-containing NAPE species were predominant over 18:1-containing species in heart, liver, and jejunum. Our results show that both molecular species composition of NAPE, NAE and Gp-NAE and their dependencies on Napepld are different among the peripheral tissues, suggesting that each tissue has distinct metabolic pathways and these NAE-containing lipids play tissue-specific roles.","null","null","2017-12-01","The Journal of Biochemistry","The Journal of Biochemistry","Vol.162","No.6","449","458","eng","true","null","scientific_journal","null","null","10.1093/jb/mvx054","1756-2651","null","null","null","null","null"
"Distribution of glycosylinositol phosphoceramide-specific phospholipase D activity in plants","Distribution of glycosylinositol phosphoceramide-specific phospholipase D activity in plants","Kida Takashi, Itoh Aoi, Kimura Akari, Matsuoka Hisatsugu, Imai Hiroyuki, Kentaro Kogure, Akira Tokumura, Tamotsu Tanaka","Kida Takashi, Itoh Aoi, Kimura Akari, Matsuoka Hisatsugu, Imai Hiroyuki, Kentaro Kogure, Akira Tokumura, Tamotsu Tanaka","null","Previously, we detected an unknown sphingophospholipid in cabbage leaves and identified it as phytoceramide-1-phosphate (PC1P). We also found an enzyme activity that produces PC1P by glycosylinositol phosphoceramide (GIPC)-specific hydrolysis in cabbage leaves. To characterize the GIPC-specific phospholipase D (GIPC-PLD) activity, we investigated distributions of GIPC-PLD activity in 25 tissues of 10 plants. In most plants, the GIPC-PLD activity was the highest in roots. Young leaves of cabbage and Welsh onion had higher activities than corresponding aged outer leaves. The GIPC-PLD activities in leaves, stems and roots of mung bean were higher in the sprouting stage than in more mature stages. We also examined the distribution of substrate GIPC and product PC1P and found that GIPC was ubiquitously distributed at 50-280 nmol/g (wet wt) in tissues of plants, whereas PC1P was detectable (3-60 nmol/g wet wt.) only in tissues showing considerable GIPC-PLD activity. These results suggest a possibility that GIPC-PLD activity is involved in plant growth.","Previously, we detected an unknown sphingophospholipid in cabbage leaves and identified it as phytoceramide-1-phosphate (PC1P). We also found an enzyme activity that produces PC1P by glycosylinositol phosphoceramide (GIPC)-specific hydrolysis in cabbage leaves. To characterize the GIPC-specific phospholipase D (GIPC-PLD) activity, we investigated distributions of GIPC-PLD activity in 25 tissues of 10 plants. In most plants, the GIPC-PLD activity was the highest in roots. Young leaves of cabbage and Welsh onion had higher activities than corresponding aged outer leaves. The GIPC-PLD activities in leaves, stems and roots of mung bean were higher in the sprouting stage than in more mature stages. We also examined the distribution of substrate GIPC and product PC1P and found that GIPC was ubiquitously distributed at 50-280 nmol/g (wet wt) in tissues of plants, whereas PC1P was detectable (3-60 nmol/g wet wt.) only in tissues showing considerable GIPC-PLD activity. These results suggest a possibility that GIPC-PLD activity is involved in plant growth.","null","null","2017-02-01","The Journal of Biochemistry","The Journal of Biochemistry","Vol.161","No.2","187","195","eng","true","null","scientific_journal","null","null","10.1093/jb/mvw060","1756-2651","null","null","null","null","null"
"Concentrated phosphatidic acid in cereal brans as potential protective agents against indomethacin-induced stomach ulcer.","Concentrated phosphatidic acid in cereal brans as potential protective agents against indomethacin-induced stomach ulcer.","S Afroz, Teru Ikoma, Ayano Yagi, Kentaro Kogure, Akira Tokumura, Tamotsu Tanaka","S Afroz, Teru Ikoma, Ayano Yagi, Kentaro Kogure, Akira Tokumura, Tamotsu Tanaka","null","One of complications associated with long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs) is peptic ulcer. Recently, we found that orally administered phosphatidic acid (PA) ameliorated aspirin-induced stomach lesions in mice. In this study, we identified PA-rich food sources and examined the effects of the food materials on indomethacin-induced stomach ulcer. Among examined, buckwheat (Fagopyrum esculentum) bran contained the highest level of PA (188 mg/100 g). PA was the richest phospholipid (25%) in the lipid fraction of the buckwheat bran. Administration of the lipid extracts of buckwheat bran significantly ameliorated indomethacin-induced stomach lesions in mice. In contrast, wheat (Triticum durum) bran lipids (PA, 4%) and soybean (Glycine max) lipids (PA, 3%) were not associated with ameliorative effects. These results indicated that PA-rich lipids can be used as an effective supplement for prevention of NSAID-induced stomach ulcer.","One of complications associated with long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs) is peptic ulcer. Recently, we found that orally administered phosphatidic acid (PA) ameliorated aspirin-induced stomach lesions in mice. In this study, we identified PA-rich food sources and examined the effects of the food materials on indomethacin-induced stomach ulcer. Among examined, buckwheat (Fagopyrum esculentum) bran contained the highest level of PA (188 mg/100 g). PA was the richest phospholipid (25%) in the lipid fraction of the buckwheat bran. Administration of the lipid extracts of buckwheat bran significantly ameliorated indomethacin-induced stomach lesions in mice. In contrast, wheat (Triticum durum) bran lipids (PA, 4%) and soybean (Glycine max) lipids (PA, 3%) were not associated with ameliorative effects. These results indicated that PA-rich lipids can be used as an effective supplement for prevention of NSAID-induced stomach ulcer.","null","null","2016-12","Journal of Agricultural and Food Chemistry","Journal of Agricultural and Food Chemistry","Vol.64","No.37","6950","6957","eng","true","null","scientific_journal","null","null","10.1021/acs.jafc.6b02884","1520-5118","null","null","null","null","null"
"Calcium-dependent generation of N-acylethanolamines and lysophosphatidic acids by glycerophosphodiesterase GDE7.","Calcium-dependent generation of N-acylethanolamines and lysophosphatidic acids by glycerophosphodiesterase GDE7.","Iffat Sonia Ara Rahman, Kazuhito Tsuboi, Zahir Hussain, Ryouhei Yamashita, Yoko Okamoto, Toru Uyama, Naoshi Yamazaki, Tamotsu Tanaka, Akira Tokumura, Natsuo Ueda","Iffat Sonia Ara Rahman, Kazuhito Tsuboi, Zahir Hussain, Ryouhei Yamashita, Yoko Okamoto, Toru Uyama, Naoshi Yamazaki, Tamotsu Tanaka, Akira Tokumura, Natsuo Ueda","null","null","null","null","null","2016-12","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1861","No.12 pt A","1881","1892","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2016.09.008","1388-1981","null","null","null","null","null"
"The novel functional nucleic acid iRed effectively regulates target genes following cytoplasmic delivery by faint electric treatment","The novel functional nucleic acid iRed effectively regulates target genes following cytoplasmic delivery by faint electric treatment","Mahadi Hasan, Noriko Saito-Tarashima, Koki Fujikawa, Takashi Ohgita, Susumu Hama, Tamotsu Tanaka, Hiroyuki Saito, Noriaki Minakawa, Kentaro Kogure","Mahadi Hasan, Noriko Saito-Tarashima, Koki Fujikawa, Takashi Ohgita, Susumu Hama, Tamotsu Tanaka, Hiroyuki Saito, Noriaki Minakawa, Kentaro Kogure","null","An intelligent shRNA expression device (iRed) contains the minimum essential components needed for shRNA production in cells, and could be a novel tool to regulate target genes. However, general delivery carriers consisting of cationic polymers/lipids could impede function of a newly generated shRNA via electrostatic interaction in the cytoplasm. Recently, we found that faint electric treatment (fET) of cells enhanced delivery of siRNA and functional nucleic acids into the cytoplasm in the absence of delivery carriers. Here, we examined fET of cells stably expressing luciferase in the presence of iRed encoding anti-luciferase shRNA. Transfection of lipofectamine 2000 (LFN)/iRed lipoplexes showed an RNAi effect, but fET-mediated iRed transfection did not, likely because of the endosomal localization of iRed after delivery. However, fET in the presence of lysosomotropic agent chloroquine significantly improved the RNAi effect of iRed/fET to levels that were higher than those for the LFN/iRed lipoplexes. Furthermore, the amount of lipid droplets in adipocytes significantly decreased following fET with iRed against resistin in the presence of chloroquine. Thus, iRed could be a useful tool to regulate target genes following fET-mediated cytoplasmic delivery with endosomal escape devices.","An intelligent shRNA expression device (iRed) contains the minimum essential components needed for shRNA production in cells, and could be a novel tool to regulate target genes. However, general delivery carriers consisting of cationic polymers/lipids could impede function of a newly generated shRNA via electrostatic interaction in the cytoplasm. Recently, we found that faint electric treatment (fET) of cells enhanced delivery of siRNA and functional nucleic acids into the cytoplasm in the absence of delivery carriers. Here, we examined fET of cells stably expressing luciferase in the presence of iRed encoding anti-luciferase shRNA. Transfection of lipofectamine 2000 (LFN)/iRed lipoplexes showed an RNAi effect, but fET-mediated iRed transfection did not, likely because of the endosomal localization of iRed after delivery. However, fET in the presence of lysosomotropic agent chloroquine significantly improved the RNAi effect of iRed/fET to levels that were higher than those for the LFN/iRed lipoplexes. Furthermore, the amount of lipid droplets in adipocytes significantly decreased following fET with iRed against resistin in the presence of chloroquine. Thus, iRed could be a useful tool to regulate target genes following fET-mediated cytoplasmic delivery with endosomal escape devices.","null","null","2016-08-04","Science and Technology of Advanced Materials","Science and Technology of Advanced Materials","Vol.17","No.17","554","562","eng","true","null","scientific_journal","null","null","10.1080/14686996.2016.1221726","1468-6996","null","null","null","null","null"
"Preferable existence of polyunsaturated lysophosphatidic acids in human follicular fluid from patients programmed with in vitro fertilization.","Preferable existence of polyunsaturated lysophosphatidic acids in human follicular fluid from patients programmed with in vitro fertilization.","Junpei Yamamoto, Midori Omura, Koichiro Tuchiya, Mayumi Hidaka, Akira Kuwahara, Minoru Irahara, Tamotsu Tanaka, Akira Tokumura","Junpei Yamamoto, Midori Omura, Koichiro Tuchiya, Mayumi Hidaka, Akira Kuwahara, Minoru Irahara, Tamotsu Tanaka, Akira Tokumura","null","Lysophosphatidic acid (LPA) exerts diverse physiological effects on various types of animal cells, including reproductive cells, through its binding to six LPA receptors. We previously found that LPA promoted maturation of the nucleus and cytoplasm of mouse and hamster oocytes surrounded by cumulus cells in vitro. Using gas-liquid chromatography, we previously reported detection of several species of LPA by analyzing the fatty acid methyl esters derived from thin layer chromatography-purified LPA in lipid extract from incubated follicular fluids programmed with in vitro fertilization. In this study using liquid chromatography- tandem mass spectrometry, we directly detected high levels of linoleoyl, arachidonoyl, and docosahexaenoyl LPAs in human follicular fluid. This unique molecular species composition of LPA was suggested to be due to a balance between the low LPA-degrading activity and high LPA-producing activity of autotaxin in human follicular fluid. Our results suggest that polyunsaturated LPAs produced by autotaxin in human follicular fluid exert unknown physiological effects on cumulus cells.","Lysophosphatidic acid (LPA) exerts diverse physiological effects on various types of animal cells, including reproductive cells, through its binding to six LPA receptors. We previously found that LPA promoted maturation of the nucleus and cytoplasm of mouse and hamster oocytes surrounded by cumulus cells in vitro. Using gas-liquid chromatography, we previously reported detection of several species of LPA by analyzing the fatty acid methyl esters derived from thin layer chromatography-purified LPA in lipid extract from incubated follicular fluids programmed with in vitro fertilization. In this study using liquid chromatography- tandem mass spectrometry, we directly detected high levels of linoleoyl, arachidonoyl, and docosahexaenoyl LPAs in human follicular fluid. This unique molecular species composition of LPA was suggested to be due to a balance between the low LPA-degrading activity and high LPA-producing activity of autotaxin in human follicular fluid. Our results suggest that polyunsaturated LPAs produced by autotaxin in human follicular fluid exert unknown physiological effects on cumulus cells.","null","null","2016-07-12","Prostaglandins & Other Lipid Mediators","Prostaglandins & Other Lipid Mediators","Vol.126","null","16","23","eng","true","null","scientific_journal","null","null","10.1016/j.prostaglandins.2016.07.008","1098-8823","null","null","null","null","null"
"Reduced rat plasma lysophosphatidylglycerol or lysophosphatidic acid level as a biomarker of aristolochic acid-induced renal and adipose dysfunctions.","Reduced rat plasma lysophosphatidylglycerol or lysophosphatidic acid level as a biomarker of aristolochic acid-induced renal and adipose dysfunctions.","Toshihiko Tsutsumi, Yoko Okamoto, Syougo Yamakawa, Cheng Bingjun, Akira Ishihara, Tamotsu Tanaka, Akira Tokumura","Toshihiko Tsutsumi, Yoko Okamoto, Syougo Yamakawa, Cheng Bingjun, Akira Ishihara, Tamotsu Tanaka, Akira Tokumura","null","Food products and diet pills containing aristolochic acid (AA) are responsible for a rapid progression of nephropathy associated with reduced body weight in human beings. In this study, we investigated the relationship of dietary NaCl and lysophospholipid (LPL) plasma levels to body weight gain in AA-treated rats. Male rats receiving a salt-deficient chow, normal salt chow or high salt chow were injected intraperitoneally daily with AA for 15days. Body weight, visceral fat mass, food intake, levels of LPL in plasma and its synthesized enzyme were investigated. Body weight gain, visceral fat mass and daily food intake were smaller in AA-treated rats than those of control rats, regardless of dietary salt concentration. AA treatment decreased plasma levels of major lysophosphatidic acid (LPA) molecular species in rats fed the normal or high-salt chow but not the salt-deficient chow, whereas both the plasma lysophospholipase D activity and kidney mRNA level of autotaxin of AA-treated rats fed chow with defined salt concentrations were lower than those of control rats. Plasma levels of major molecular species of lysophosphatidylglycerol (LPG) in AA-treated rat groups fed chow with defined salt concentrations were lower than those of control rats. Plasma levels of LPG and LPA seem to be relevant to the reduced body weight gain and fat mass due to AA treatment.","Food products and diet pills containing aristolochic acid (AA) are responsible for a rapid progression of nephropathy associated with reduced body weight in human beings. In this study, we investigated the relationship of dietary NaCl and lysophospholipid (LPL) plasma levels to body weight gain in AA-treated rats. Male rats receiving a salt-deficient chow, normal salt chow or high salt chow were injected intraperitoneally daily with AA for 15days. Body weight, visceral fat mass, food intake, levels of LPL in plasma and its synthesized enzyme were investigated. Body weight gain, visceral fat mass and daily food intake were smaller in AA-treated rats than those of control rats, regardless of dietary salt concentration. AA treatment decreased plasma levels of major lysophosphatidic acid (LPA) molecular species in rats fed the normal or high-salt chow but not the salt-deficient chow, whereas both the plasma lysophospholipase D activity and kidney mRNA level of autotaxin of AA-treated rats fed chow with defined salt concentrations were lower than those of control rats. Plasma levels of major molecular species of lysophosphatidylglycerol (LPG) in AA-treated rat groups fed chow with defined salt concentrations were lower than those of control rats. Plasma levels of LPG and LPA seem to be relevant to the reduced body weight gain and fat mass due to AA treatment.","null","null","2016-06-04","Life Sciences","Life Sciences","Vol.157","null","208","216","eng","true","null","scientific_journal","null","null","10.1016/j.lfs.2016.06.003","1879-0631","null","null","null","null","null"
"食品に含まれるグリコシルイノシトールホスホセラミドおよびフィトセラミド-1-リン酸","食品に含まれるグリコシルイノシトールホスホセラミドおよびフィトセラミド-1-リン酸","喜田 孝史, 木村 朱里, 伊藤 葵, 山下 量平, 小暮 健太朗, 德村 彰, 田中 保","喜田 孝史, 木村 朱里, 伊藤 葵, 山下 量平, Kentaro Kogure, Akira Tokumura, Tamotsu Tanaka","null","null","null","null","null","2016","脂質栄養学","Journal of Lipid Nutrition","Vol.25","null","75","85","jpn","true","null","scientific_journal","null","null","10.4010/jln.25.75","1343-4594","null","null","null","null","null"
"Analysis of molecular species profiles of ceramide-1-phosphate and sphingomyelin using MALDI-TOF mass spectrometry","Analysis of molecular species profiles of ceramide-1-phosphate and sphingomyelin using MALDI-TOF mass spectrometry","Ryouhei Yamashita, Yumika Tabata, Erina Iga, Michiyasu Nakao, Shigeki Sano, Kentaro Kogure, Akira Tokumura, Tamotsu Tanaka","Ryouhei Yamashita, Yumika Tabata, Erina Iga, Michiyasu Nakao, Shigeki Sano, Kentaro Kogure, Akira Tokumura, Tamotsu Tanaka","null","Ceramide-1-phosphate (C1P) is a potential signaling molecule that modulates various cellular functions in animals. It has been known that C1P with different N-acyl lengths induce biological responses differently. However, molecular species profiles of the C1P in animal tissues have not been extensively examined yet. Here, we developed a method for determination of the molecular species of a C1P using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with Phos-tag, a phosphate capture molecule. The amounts of total C1P in skin, brain, liver, kidney and small intestine of mice were determined to be 344, 151, 198, 96 and 90 pmol/g wet weight, respectively. We found a C1P species having an α-hydroxypalmitoyl residue (h-C1P, 44 pmol/g wet weight) in mouse skin. The h-C1P was detected only in the skin, and not other tissues of mice. The same analysis was applied to sphingomyelin after conversion of sphingomyelin to C1P by Streptomyces chromofuscus phospholipase D. We found that molecular species profiles of sphingomyelin in skin, kidney and small intestine of mice were similar to those of C1P in corresponding tissues. In contrast, molecular species profiles of sphingomyelin in liver and brain were quite different from those of C1P in these tissues, indicating selective synthesis or degradation of C1P in these tissues. The method described here will be useful for detection of changes in molecular species profiles of C1P and sphingomyelin.","Ceramide-1-phosphate (C1P) is a potential signaling molecule that modulates various cellular functions in animals. It has been known that C1P with different N-acyl lengths induce biological responses differently. However, molecular species profiles of the C1P in animal tissues have not been extensively examined yet. Here, we developed a method for determination of the molecular species of a C1P using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with Phos-tag, a phosphate capture molecule. The amounts of total C1P in skin, brain, liver, kidney and small intestine of mice were determined to be 344, 151, 198, 96 and 90 pmol/g wet weight, respectively. We found a C1P species having an α-hydroxypalmitoyl residue (h-C1P, 44 pmol/g wet weight) in mouse skin. The h-C1P was detected only in the skin, and not other tissues of mice. The same analysis was applied to sphingomyelin after conversion of sphingomyelin to C1P by Streptomyces chromofuscus phospholipase D. We found that molecular species profiles of sphingomyelin in skin, kidney and small intestine of mice were similar to those of C1P in corresponding tissues. In contrast, molecular species profiles of sphingomyelin in liver and brain were quite different from those of C1P in these tissues, indicating selective synthesis or degradation of C1P in these tissues. The method described here will be useful for detection of changes in molecular species profiles of C1P and sphingomyelin.","null","null","2016","Lipids","Lipids","Vol.51","No.2","263","270","eng","true","null","scientific_journal","null","null","10.1007/s11745-015-4082-0","1558-9307","null","null","null","null","null"
"Reduced kidney levels of lysophosphatidic acids in rats after chronic administration of aristolochic acid: Its possible protective role in renal fibrosis","Reduced kidney levels of lysophosphatidic acids in rats after chronic administration of aristolochic acid: Its possible protective role in renal fibrosis","Toshihiko Tsutsumi, Syougo Yamakawa, Akira Ishihara, Aimi Yamamoto, Tamotsu Tanaka, Akira Tokumura","Toshihiko Tsutsumi, Syougo Yamakawa, Akira Ishihara, Aimi Yamamoto, Tamotsu Tanaka, Akira Tokumura","null","null","null","null","null","2015-02","Toxicology Reports","Toxicology Reports","Vol.2","null","121","129","eng","true","null","scientific_journal","null","null","10.1016/j.toxrep.2015.02.012","2214-7500","null","null","null","null","null"
"Potentials of the circulating pruritogenic mediator lysophosphatidic acid in development of allergic skin inflammation in mice: role of blood cell-associated lysophospholipase D activity of autotaxin.","Potentials of the circulating pruritogenic mediator lysophosphatidic acid in development of allergic skin inflammation in mice: role of blood cell-associated lysophospholipase D activity of autotaxin.","Yoshibumi Shimizu, Yoshiyuki Morikawa, Shinichi Okudaira, Shigenobu Kimoto, Tamotsu Tanaka, Junken Aoki, Akira Tokumura","Yoshibumi Shimizu, Yoshiyuki Morikawa, Shinichi Okudaira, Shigenobu Kimoto, Tamotsu Tanaka, Junken Aoki, Akira Tokumura","null","Itching and infiltration of immune cells are important hallmarks of atopic dermatitis (AD). Although various studies have focused on peripheral mediator-mediated mechanisms, systemic mediator-mediated mechanisms are also important in the pathogenesis and development of AD. Herein, we found that intradermal injection of lysophosphatidic acid (LPA), a bioactive phospholipid, induces scratching responses by Institute of Cancer Research mice through LPA1 receptor- and opioid μ receptor-mediating mechanisms, indicating its potential as a pruritogen. The circulating level of LPA in Naruto Research Institute Otsuka Atrichia mice, a systemic AD model, with severe scratching was found to be higher than that of control BALB/c mice, probably because of the increased lysophospholipase D activity of autotaxin (ATX) in the blood (mainly membrane associated) rather than in plasma (soluble). Heparan sulfate proteoglycan was shown to be involved in the association of ATX with blood cells. The sequestration of ATX protein on the blood cells by heparan sulfate proteoglycan may accelerate the transport of LPA to the local apical surface of vascular endothelium with LPA receptors, promoting the hyperpermeability of venules and the pathological uptake of immune cells, aggravating lesion progression and itching in Naruto Research Institute Otsuka Atrichia mice.","Itching and infiltration of immune cells are important hallmarks of atopic dermatitis (AD). Although various studies have focused on peripheral mediator-mediated mechanisms, systemic mediator-mediated mechanisms are also important in the pathogenesis and development of AD. Herein, we found that intradermal injection of lysophosphatidic acid (LPA), a bioactive phospholipid, induces scratching responses by Institute of Cancer Research mice through LPA1 receptor- and opioid μ receptor-mediating mechanisms, indicating its potential as a pruritogen. The circulating level of LPA in Naruto Research Institute Otsuka Atrichia mice, a systemic AD model, with severe scratching was found to be higher than that of control BALB/c mice, probably because of the increased lysophospholipase D activity of autotaxin (ATX) in the blood (mainly membrane associated) rather than in plasma (soluble). Heparan sulfate proteoglycan was shown to be involved in the association of ATX with blood cells. The sequestration of ATX protein on the blood cells by heparan sulfate proteoglycan may accelerate the transport of LPA to the local apical surface of vascular endothelium with LPA receptors, promoting the hyperpermeability of venules and the pathological uptake of immune cells, aggravating lesion progression and itching in Naruto Research Institute Otsuka Atrichia mice.","null","null","2014-05","The American Journal of Pathology","The American Journal of Pathology","Vol.184","No.5","1593","1603","eng","true","null","scientific_journal","null","null","10.1016/j.ajpath.2014.01.029","1525-2191","null","null","null","null","null"
"Metabolic conversion of C20 polymethylene-interrupted polyunsaturated fatty acids to essential fatty acids.","Metabolic conversion of C20 polymethylene-interrupted polyunsaturated fatty acids to essential fatty acids.","Tamotsu Tanaka, Sachika Uozumi, Katsuya Morito, Takashi Osumi, Akira Tokumura","Tamotsu Tanaka, Sachika Uozumi, Katsuya Morito, Takashi Osumi, Akira Tokumura","null","Polymethylene-interrupted (PMI)-polyunsaturated fatty acids (PUFA) are fatty acids present largely in gymnosperm. Sciadonic acid (SciA, 20:3 Δ-5,11,14) and juniperonic acid (JA, 20:4 Δ-5,11,14,17) are typical C20 PMI-PUFA with an isolated double bond at Δ5. Previously, we found that SciA and JA are converted to linoleic acid (LNA) and α-linolenic acid (ΑLA), respectively. The conversion process includes chain-shortening step by peroxisomal β-oxidation for elimination a double bond at Δ5, and subsequent chain-elongation step in microsomes. In this study, we examined the substrate specificity of this metabolism in rodent and human cells. Supplementation of SciA, eicosadienoic acid (EDA, 20:2 Δ-11,14) or JA to CHO-K1 cells (wild type) induced an accumulation of LNA, LNA or ALA, respectively, in cellular lipids. These changes were not observed in the peroxisomes-deficient CHO cells, indicating involvement of peroxisomes in the metabolism. Two types of human cells (MKN74 and HepG2) also converted the C20 PMI-PUFA and EDA to the respective essential fatty acids. In contrast, no chain-shortened metabolite of pinolenic acid (18:3 Δ-5,9,12) was detected in any cell lines tested. From these results, C20 PMI-PUFA and EDA, but not C18 PMI-PUFA, are suggested as being effectively converted to essential fatty acids by the fatty acid remodeling system in rodent and human cells.","Polymethylene-interrupted (PMI)-polyunsaturated fatty acids (PUFA) are fatty acids present largely in gymnosperm. Sciadonic acid (SciA, 20:3 Δ-5,11,14) and juniperonic acid (JA, 20:4 Δ-5,11,14,17) are typical C20 PMI-PUFA with an isolated double bond at Δ5. Previously, we found that SciA and JA are converted to linoleic acid (LNA) and α-linolenic acid (ΑLA), respectively. The conversion process includes chain-shortening step by peroxisomal β-oxidation for elimination a double bond at Δ5, and subsequent chain-elongation step in microsomes. In this study, we examined the substrate specificity of this metabolism in rodent and human cells. Supplementation of SciA, eicosadienoic acid (EDA, 20:2 Δ-11,14) or JA to CHO-K1 cells (wild type) induced an accumulation of LNA, LNA or ALA, respectively, in cellular lipids. These changes were not observed in the peroxisomes-deficient CHO cells, indicating involvement of peroxisomes in the metabolism. Two types of human cells (MKN74 and HepG2) also converted the C20 PMI-PUFA and EDA to the respective essential fatty acids. In contrast, no chain-shortened metabolite of pinolenic acid (18:3 Δ-5,9,12) was detected in any cell lines tested. From these results, C20 PMI-PUFA and EDA, but not C18 PMI-PUFA, are suggested as being effectively converted to essential fatty acids by the fatty acid remodeling system in rodent and human cells.","null","null","2014-05","Lipids","Lipids","Vol.49","No.5","423","429","eng","true","null","scientific_journal","null","null","10.1007/s11745-014-3896-5","1558-9307","null","null","null","null","null"
"Type 2 lysophosphatidic acid receptor in gastric surface mucous cells: Possible implication of prostaglandin E2 production","Type 2 lysophosphatidic acid receptor in gastric surface mucous cells: Possible implication of prostaglandin E2 production","Tamotsu Tanaka, Mayumi Ohmoto, Katsuya Morito, H. Kondo, Mai Urikura, Kiyoshi Satouchi, Akira Tokumura","Tamotsu Tanaka, Mayumi Ohmoto, Katsuya Morito, H. Kondo, Mai Urikura, Kiyoshi Satouchi, Akira Tokumura","null","Lysophosphatidic acid (LPA) is a lipid mediator that induces various cell responses via its specific receptors. Recently, we found that orally administered LPA and phosphatidic acid (PA) ameliorate stress- or aspirin-induced stomach injury. However, the mechanisms underlying these effects have not been elucidated yet. In this study, we examined effect of LPA on prostaglandin (PG) E2 production in MKN74 cells, a gastric cell-line expressing type 2 LPA receptor (LPA2). When the cells were treated with LPA, the level of mRNA of COX-2 but not COX-1 was upregulated. The LPA effect was abolished when the cells were pretreated with pertussis toxin (PTX), suggesting the involvement of receptor(s) coupled with Gi. Pretreatment of MKN74 cells with LPA enhanced the PGE2 production triggered by calcium ionophore A23187. Again, PTX abolished the LPA effect. Fluorescent immunohistochemistry using an antibody against LPA2 showed that surface mucous cells (pit cells) in gastric mucosa of mice express LPA2 on the apical side of the plasma membrane. These results suggest that LPA in the diet or its digestion may contribute to the epithelial integrity of stomach mucosa by enhancement of PGE2 production via activation of LPA2. © 2013 BioFactors, 2013.","Lysophosphatidic acid (LPA) is a lipid mediator that induces various cell responses via its specific receptors. Recently, we found that orally administered LPA and phosphatidic acid (PA) ameliorate stress- or aspirin-induced stomach injury. However, the mechanisms underlying these effects have not been elucidated yet. In this study, we examined effect of LPA on prostaglandin (PG) E2 production in MKN74 cells, a gastric cell-line expressing type 2 LPA receptor (LPA2). When the cells were treated with LPA, the level of mRNA of COX-2 but not COX-1 was upregulated. The LPA effect was abolished when the cells were pretreated with pertussis toxin (PTX), suggesting the involvement of receptor(s) coupled with Gi. Pretreatment of MKN74 cells with LPA enhanced the PGE2 production triggered by calcium ionophore A23187. Again, PTX abolished the LPA effect. Fluorescent immunohistochemistry using an antibody against LPA2 showed that surface mucous cells (pit cells) in gastric mucosa of mice express LPA2 on the apical side of the plasma membrane. These results suggest that LPA in the diet or its digestion may contribute to the epithelial integrity of stomach mucosa by enhancement of PGE2 production via activation of LPA2. © 2013 BioFactors, 2013.","null","null","2014-05","BioFactors","BioFactors","Vol.40","No.3","355","361","eng","true","null","scientific_journal","null","null","10.1002/biof.1147","1872-8081","null","null","null","null","null"
"Increased lysophospholipase D activity of autotaxin in sera of patients with atopic dermatitis.","Increased lysophospholipase D activity of autotaxin in sera of patients with atopic dermatitis.","Yoshibumi Shimizu, Kazutoshi Murao, Tamotsu Tanaka, Yoshiaki Kubo, Akira Tokumura","Yoshibumi Shimizu, Kazutoshi Murao, Tamotsu Tanaka, Yoshiaki Kubo, Akira Tokumura","null","null","null","null","null","2014-02-06","Journal of Dermatological Science","Journal of Dermatological Science","Vol.74","No.2","162","165","eng","true","null","scientific_journal","null","null","10.1016/j.jdermsci.2014.01.010","1873-569X","null","null","null","null","null"
"Identification of a sphingolipid-specific phospholipase D activity associated with the generation of phytoceramide-1-phosphate in cabbage leaves","Identification of a sphingolipid-specific phospholipase D activity associated with the generation of phytoceramide-1-phosphate in cabbage leaves","Tamotsu Tanaka, T. Kida, H. Imai, J. Morishige, R. Yamashita, H. Matsuoka, S. Uozumi, K. Satouchi, M. Nagano, Akira Tokumura","Tamotsu Tanaka, T. Kida, H. Imai, J. Morishige, R. Yamashita, H. Matsuoka, S. Uozumi, K. Satouchi, M. Nagano, Akira Tokumura","null","The structure and biosynthetic route for an unidentified lipid (lipid X) detected by TLC of cabbage (Brassica oleracea) lipids was determined. Lipid X is a phospholipid that is resistant to mild alkali and detectable by MALDI-TOF MS as an adduct with Phos-tag, a phosphate-capture zinc complex. Various α-hydroxy fatty acids (16:0, 22:0, 24:0 and 24:1) were detected by GC-MS of fatty acid methyl esters prepared from lipid X. The deacyl derivative of lipid X was determined to be 4-hydroxysphingenine (dehydrophytosphingosine)-1-phosphate by MALDI-TOF MS with Phos-tag. From these results, lipid X was determined to be phytoceramide-1-phosphate (PC1P) with an α-hydroxy fatty acid. When cabbage homogenates were incubated, PC1P was formed, with a concomitant decrease in the amount of glycosylinositol phosphoceramide (GIPC). The formation of PC1P from GIPC was confirmed by treatment of purified cabbage GIPC with a membrane fraction of cabbage homogenates. Using a partially purified enzyme fraction, we found that the enzyme hydrolyzes GIPC specifically, but not glycerophospholipids and sphingomyelin. Arabidopsis thaliana also had this enzyme activity. From these results, we conclude that a previously uncharacterized phospholipase D activity that specifically hydrolyzes GIPC produces PC1P in brassicaceous plants.","The structure and biosynthetic route for an unidentified lipid (lipid X) detected by TLC of cabbage (Brassica oleracea) lipids was determined. Lipid X is a phospholipid that is resistant to mild alkali and detectable by MALDI-TOF MS as an adduct with Phos-tag, a phosphate-capture zinc complex. Various α-hydroxy fatty acids (16:0, 22:0, 24:0 and 24:1) were detected by GC-MS of fatty acid methyl esters prepared from lipid X. The deacyl derivative of lipid X was determined to be 4-hydroxysphingenine (dehydrophytosphingosine)-1-phosphate by MALDI-TOF MS with Phos-tag. From these results, lipid X was determined to be phytoceramide-1-phosphate (PC1P) with an α-hydroxy fatty acid. When cabbage homogenates were incubated, PC1P was formed, with a concomitant decrease in the amount of glycosylinositol phosphoceramide (GIPC). The formation of PC1P from GIPC was confirmed by treatment of purified cabbage GIPC with a membrane fraction of cabbage homogenates. Using a partially purified enzyme fraction, we found that the enzyme hydrolyzes GIPC specifically, but not glycerophospholipids and sphingomyelin. Arabidopsis thaliana also had this enzyme activity. From these results, we conclude that a previously uncharacterized phospholipase D activity that specifically hydrolyzes GIPC produces PC1P in brassicaceous plants.","null","null","2013-07-05","The FEBS Journal","The FEBS Journal","Vol.280","No.16","3797","3809","eng","true","null","scientific_journal","null","null","10.1111/febs.12374","1742-4658","null","null","null","null","null"
"Orally administered phosphatidic acids and lysophosphatidic acids ameliorate aspirin-induced stomach mucosal injury in mice","Orally administered phosphatidic acids and lysophosphatidic acids ameliorate aspirin-induced stomach mucosal injury in mice","Tamotsu Tanaka, K. Morito, M. Kinoshita, M. Ohmoto, M. Urikura, K. Satouchi, Akira Tokumura","Tamotsu Tanaka, K. Morito, M. Kinoshita, M. Ohmoto, M. Urikura, K. Satouchi, Akira Tokumura","null","Recent investigations revealed that lysophosphatidic acid (LPA), a phospholipid with a growth factor-like activity, plays an important role in the integrity of the gastrointestinal tract epithelium. This paper attempts to clarify the effect of orally administered phosphatidic acid (PA) and LPA on aspirin-induced gastric lesions in mice. Phospholipids, a free fatty acid, a diacylglycerol and a triglyceride at 1 mM (5.7 μmol/kg body weight) or 0.1 mM were orally administered to mice 0.5 h before oral administration of aspirin (1.7 mmol/kg). The total length of lesions formed on the stomach wall was measured as a lesion index. Formation of LPA from PA in the mouse stomach was examined by in vitro (in stomach lavage fluid), ex vivo (in an isolated stomach) and in vivo (in the stomach of a living mouse) examinations of phospholipase activity. Palmitic acid, dioleoyl-glycerol, olive oil and lysophosphatidylcholine did not affect the aspirin-induced lesions. In contrast, phosphatidylcholine (1 mM), LPA (1 mM) and PA (0.1, 1 mM) significantly reduced the lesion index. Evidence for formation of LPA from PA in the stomach by gastric phospholipase A2 was obtained by in vitro, ex vivo and in vivo experiments. An LPA-specific receptor, LPA2, was found to be localized on the gastric surface-lining cells of mice. Pretreatment with PA-rich diets may prevent nonsteroidal anti-inflammatory drug-induced stomach ulcers.","Recent investigations revealed that lysophosphatidic acid (LPA), a phospholipid with a growth factor-like activity, plays an important role in the integrity of the gastrointestinal tract epithelium. This paper attempts to clarify the effect of orally administered phosphatidic acid (PA) and LPA on aspirin-induced gastric lesions in mice. Phospholipids, a free fatty acid, a diacylglycerol and a triglyceride at 1 mM (5.7 μmol/kg body weight) or 0.1 mM were orally administered to mice 0.5 h before oral administration of aspirin (1.7 mmol/kg). The total length of lesions formed on the stomach wall was measured as a lesion index. Formation of LPA from PA in the mouse stomach was examined by in vitro (in stomach lavage fluid), ex vivo (in an isolated stomach) and in vivo (in the stomach of a living mouse) examinations of phospholipase activity. Palmitic acid, dioleoyl-glycerol, olive oil and lysophosphatidylcholine did not affect the aspirin-induced lesions. In contrast, phosphatidylcholine (1 mM), LPA (1 mM) and PA (0.1, 1 mM) significantly reduced the lesion index. Evidence for formation of LPA from PA in the stomach by gastric phospholipase A2 was obtained by in vitro, ex vivo and in vivo experiments. An LPA-specific receptor, LPA2, was found to be localized on the gastric surface-lining cells of mice. Pretreatment with PA-rich diets may prevent nonsteroidal anti-inflammatory drug-induced stomach ulcers.","null","null","2013","Digestive Diseases and Sciences","Digestive Diseases and Sciences","Vol.58","No.4","950","958","eng","true","null","scientific_journal","null","null","10.1007/s10620-012-2475-y","1573-2568","null","null","null","null","null"
"Phosphatidic acid production in the processing of cabbage leaves","Phosphatidic acid production in the processing of cabbage leaves","Mai Urikura, Jun-ichi Morishige, Tamotsu Tanaka, Kiyoshi Satouchi","Mai Urikura, Jun-ichi Morishige, Tamotsu Tanaka, Kiyoshi Satouchi","null","Lysophosphatidic acid (LPA) is a lipid mediator involved in various physiological responses, including wound healing. Evidence of the antiulcer activity of LPA has been reported, and soybean LPA at a concentration of 10 μM is effective in reducing stress-induced gastric ulcer. Because LPA can be formed from phosphatidic acid (PA) by digestive phospholipase A2, dietary PA can be considered a potential antiulcer phospholipid. In this study, PA production in cut processing of cabbage leaves was examined. The amounts of PA in sliced, minced, and homogenized cabbage leaves were 107 ± 5, 134 ± 19, and 286 ± 29 nmol PA/g (wet weight), respectively, all being significantly higher than the amount of PA found in intact leaves. Mixing mayonnaise with sliced cabbage dramatically increased the PA content (1586 ± 393 nmol/3 g), indicating phospholipase D activity leaked raw cabbage produced PA. These results indicate that fine cutting raw cabbage leaves and mixing them with foods rich in phospholipids resulted in an abundant production of PA.","Lysophosphatidic acid (LPA) is a lipid mediator involved in various physiological responses, including wound healing. Evidence of the antiulcer activity of LPA has been reported, and soybean LPA at a concentration of 10 μM is effective in reducing stress-induced gastric ulcer. Because LPA can be formed from phosphatidic acid (PA) by digestive phospholipase A2, dietary PA can be considered a potential antiulcer phospholipid. In this study, PA production in cut processing of cabbage leaves was examined. The amounts of PA in sliced, minced, and homogenized cabbage leaves were 107 ± 5, 134 ± 19, and 286 ± 29 nmol PA/g (wet weight), respectively, all being significantly higher than the amount of PA found in intact leaves. Mixing mayonnaise with sliced cabbage dramatically increased the PA content (1586 ± 393 nmol/3 g), indicating phospholipase D activity leaked raw cabbage produced PA. These results indicate that fine cutting raw cabbage leaves and mixing them with foods rich in phospholipids resulted in an abundant production of PA.","null","null","2012-11-01","Journal of Agricultural and Food Chemistry","Journal of Agricultural and Food Chemistry","Vol.60","No.45","11359","11365","eng","true","null","scientific_journal","null","null","10.1021/jf303515z","1520-5118","null","null","null","null","null"
"Quantification of phosphatidic acid in foodstuffs using TLC-imaging technique","Quantification of phosphatidic acid in foodstuffs using TLC-imaging technique","Tamotsu Tanaka, Ayaka Kassai, Mayumi Ohmoto, Katsuya Morito, Yoshiki Kashiwada, Yoshihisa Takaishi, Mai Urikura, Jun-ichi Morishige, Kiyoshi Satouchi, Akira Tokumura","Tamotsu Tanaka, Ayaka Kassai, Mayumi Ohmoto, Katsuya Morito, Yoshiki Kashiwada, Yoshihisa Takaishi, Mai Urikura, Jun-ichi Morishige, Kiyoshi Satouchi, Akira Tokumura","null","Apical application of lysophosphatidic acid (LPA), a growth-factor-like phospholipid, was shown to prevent or restore gastrointestinal (GI) disorders, such as diarrhea and stomach ulcer, in experimental animals. Because LPA is formed from phosphatidic acid (PA) by the activity of digestive phospholipase A(2), PA is a potential component for dietary treatment of such GI disorders. Here, we quantified PA contained in 38 foodstuffs and 3 herbs by a thin-layer-chromatography-imaging technique. Vegetables belonging to Brassicaceae, such as cabbage leaves (700 nmol/g of wet weight) and Japanese radish leaves (570 nmol/g), contained higher amounts of PA than other foodstuffs. Amounts of PA in fruits, cereals, and starchy root vegetables were below 300 nmol/g. Animal foodstuffs contained low amounts of PA (<60 nmol/g). Interestingly, leaves of Mallotus japonicas, a Japanese edible herb used for treatment of stomach ulcer, had the highest PA (1410 nmol/g) among those examined. The data shown here will be useful for the development of dietary treatment for a damaged GI tract.","Apical application of lysophosphatidic acid (LPA), a growth-factor-like phospholipid, was shown to prevent or restore gastrointestinal (GI) disorders, such as diarrhea and stomach ulcer, in experimental animals. Because LPA is formed from phosphatidic acid (PA) by the activity of digestive phospholipase A(2), PA is a potential component for dietary treatment of such GI disorders. Here, we quantified PA contained in 38 foodstuffs and 3 herbs by a thin-layer-chromatography-imaging technique. Vegetables belonging to Brassicaceae, such as cabbage leaves (700 nmol/g of wet weight) and Japanese radish leaves (570 nmol/g), contained higher amounts of PA than other foodstuffs. Amounts of PA in fruits, cereals, and starchy root vegetables were below 300 nmol/g. Animal foodstuffs contained low amounts of PA (<60 nmol/g). Interestingly, leaves of Mallotus japonicas, a Japanese edible herb used for treatment of stomach ulcer, had the highest PA (1410 nmol/g) among those examined. The data shown here will be useful for the development of dietary treatment for a damaged GI tract.","null","null","2012-04-16","Journal of Agricultural and Food Chemistry","Journal of Agricultural and Food Chemistry","Vol.60","No.16","4156","4161","eng","true","null","scientific_journal","null","null","10.1021/jf300147y","1520-5118","null","null","null","null","null"
"Phos-tagを用いた活性リン脂質の質量分析","Mass spectrometric analysis of phospholipids with phosphate monoester residue using Phos-tag","田中 保, 盛重 純一, 瓜倉 真衣, 德村 彰, 里内 清","Tamotsu Tanaka, 盛重 純一, 瓜倉 真衣, Akira Tokumura, 里内 清","null","Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) are growth factor-like bioactive lipids having a phosphate monoester residue. Phos-tag can bind to them and be used both for purification and quantification. In a two-phase solvent system consisting of chloroform/methanol/water, addition of Phos-tag move LPA and S1P from a hydrophilic phase to a hydrophobic phase in the form of their Phos-tag complexes. Using this property, we developed a method for purification of LPA and S1P in biological materials by the phase separation technique. Advantages of use of Phos-tag for detection of LPA and S1P in MALDI-TOF MS are an increase in ionization efficiency and detection as a single-ion form. Homologues of LPA and S1P in natural samples can be quantified by MALDI-TOF MS by using internal standards.
","Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) are growth factor-like bioactive lipids having a phosphate monoester residue. Phos-tag can bind to them and be used both for purification and quantification. In a two-phase solvent system consisting of chloroform/methanol/water, addition of Phos-tag move LPA and S1P from a hydrophilic phase to a hydrophobic phase in the form of their Phos-tag complexes. Using this property, we developed a method for purification of LPA and S1P in biological materials by the phase separation technique. Advantages of use of Phos-tag for detection of LPA and S1P in MALDI-TOF MS are an increase in ionization efficiency and detection as a single-ion form. Homologues of LPA and S1P in natural samples can be quantified by MALDI-TOF MS by using internal standards.
","null","null","2012-01","生物物理化学","SEIBUTSU BUTSURI KAGAKU","Vol.56","null","s37","42","jpn","true","true","scientific_journal","null","null","10.2198/sbk.56.s37","0031-9082","null","http://id.ndl.go.jp/bib/023553759","null","null","null"
"Alterations of plasma levels of lysophosphatidic acid in response to fasting of rats","Alterations of plasma levels of lysophosphatidic acid in response to fasting of rats","Masaki Ino, Yoshibumi Shimizu, Tamotsu Tanaka, Akira Tokumura","Masaki Ino, Yoshibumi Shimizu, Tamotsu Tanaka, Akira Tokumura","null","The aim of this study was to investigate the effect of fasting on in vivo plasma levels of lysophosphatidic acid (LPA), a physiologically important lysophospholipid mediator. We assayed to measure activities of an LPA-producing enzyme (lysophospholipase D) and LPA-degrading enzyme activities (lysophspholipase A, lipid phosphate phosphatase) in rat plasma or blood, by measuring choline, fatty acid and inorganic phosphate, respectively. Both LPA and its precursor lysophosphatidylcholine (LPC) were quantified by liquid chromatography-tandem mass spectrometry. Fasting of rats for 24 h decreased plasma concentrations of oleoyl-, linoleoyl-, arachidonoyl- and docosahexaenoyl-LPAs, but not palmitoyl- and stearoyl-LPAs, possibly due to decreased levels of corresponding LPCs in the plasma and elevated lipid phosphate phosphatase activity for LPAs in the blood. Our results indicate that the in vivo circulating levels of LPAs in rats are affected by fasting.","The aim of this study was to investigate the effect of fasting on in vivo plasma levels of lysophosphatidic acid (LPA), a physiologically important lysophospholipid mediator. We assayed to measure activities of an LPA-producing enzyme (lysophospholipase D) and LPA-degrading enzyme activities (lysophspholipase A, lipid phosphate phosphatase) in rat plasma or blood, by measuring choline, fatty acid and inorganic phosphate, respectively. Both LPA and its precursor lysophosphatidylcholine (LPC) were quantified by liquid chromatography-tandem mass spectrometry. Fasting of rats for 24 h decreased plasma concentrations of oleoyl-, linoleoyl-, arachidonoyl- and docosahexaenoyl-LPAs, but not palmitoyl- and stearoyl-LPAs, possibly due to decreased levels of corresponding LPCs in the plasma and elevated lipid phosphate phosphatase activity for LPAs in the blood. Our results indicate that the in vivo circulating levels of LPAs in rats are affected by fasting.","null","null","2012","Biological & Pharmaceutical Bulletin","Biological & Pharmaceutical Bulletin","Vol.35","No.11","2059","2063","eng","true","null","scientific_journal","null","null","10.1248/bpb.b12-00497","1347-5215","null","null","null","null","null"
"Intragastrically administrered lysophospatidic acid protect against gastric ulcer in rats under water-immersion restraint stress","Intragastrically administrered lysophospatidic acid protect against gastric ulcer in rats under water-immersion restraint stress","MIka Adachi, Gou Horiuchi, Natsuki Ikematsu, Tamotsu Tanaka, Shigeki Sano, Kenji Fukuzawa, Akira Tokumura","MIka Adachi, Gou Horiuchi, Natsuki Ikematsu, Tamotsu Tanaka, Shigeki Sano, Kenji Fukuzawa, Akira Tokumura","null","Lysophosphatidic acid exerts important physiological effects on many types of animal cells through its specific binding to several G protein-coupled receptors. In particular, its potent wound-healing effect has attracted much attention. To determine whether lysophosphatidic acids in a foodstuff and Chinese medicine are effective in protecting against gastric ulcer, we subjected rats to water-immersion restraint stress. Three direct administrations of a solution of lysophosphatidic acid with a C18 fatty acyl group to the rat stomach in a concentration range of 0.001-0.1 mM resulted in a significant reduction in the number of gastric ulcers induced during water-immersion restraint stress, and the potencies were as follows: linoleoyl species=α-linolenoyl species>oleoyl species. Intragastric administrations of a solution of highly purified lysophosphatidic acid from soybean lecithin significantly protected against the stress-induced gastric ulcers at lower concentrations than partially purified lysophosphatidic acid from soybean lecithin did. In addition, administration of a decocted solution of antyu-san, and lysophosphatidic acid-rich Chinese medicine, to the stomach was more effective in protecting against stress-induced ulcer than decoctations of antyu-san lacking the corydalis tuber component that is rich in lysophosphatidic acid. These results clearly show that lysophosphatidic acid is the effective component of soybean lecithin and antyu-san in protection against stress-induced gastric ulcer in the rat model, and suggest that daily intake of lysophosphatidic acid-rich foods or Chinese medicines may be beneficial for prevention of stress-induced gastric ulcer in human subjects.","Lysophosphatidic acid exerts important physiological effects on many types of animal cells through its specific binding to several G protein-coupled receptors. In particular, its potent wound-healing effect has attracted much attention. To determine whether lysophosphatidic acids in a foodstuff and Chinese medicine are effective in protecting against gastric ulcer, we subjected rats to water-immersion restraint stress. Three direct administrations of a solution of lysophosphatidic acid with a C18 fatty acyl group to the rat stomach in a concentration range of 0.001-0.1 mM resulted in a significant reduction in the number of gastric ulcers induced during water-immersion restraint stress, and the potencies were as follows: linoleoyl species=α-linolenoyl species>oleoyl species. Intragastric administrations of a solution of highly purified lysophosphatidic acid from soybean lecithin significantly protected against the stress-induced gastric ulcers at lower concentrations than partially purified lysophosphatidic acid from soybean lecithin did. In addition, administration of a decocted solution of antyu-san, and lysophosphatidic acid-rich Chinese medicine, to the stomach was more effective in protecting against stress-induced ulcer than decoctations of antyu-san lacking the corydalis tuber component that is rich in lysophosphatidic acid. These results clearly show that lysophosphatidic acid is the effective component of soybean lecithin and antyu-san in protection against stress-induced gastric ulcer in the rat model, and suggest that daily intake of lysophosphatidic acid-rich foods or Chinese medicines may be beneficial for prevention of stress-induced gastric ulcer in human subjects.","null","null","2011-02-06","Digestive Diseases and Sciences","Digestive Diseases and Sciences","Vol.56","No.8","2252","2261","eng","true","null","scientific_journal","null","null","10.1007/s10620-011-1595-0","1573-2568","null","null","null","null","null"
"Evaluation of inhibitory actions of flavonols and related substances on lysophospholipase d activity of serum autotaxin by a convenient assay using a chromogenic substrate.","Evaluation of inhibitory actions of flavonols and related substances on lysophospholipase d activity of serum autotaxin by a convenient assay using a chromogenic substrate.","Kaori Ueda, Masanori Yoshihara, Michiyasu Nakao, Tamotsu Tanaka, Shigeki Sano, Kenji Fukuzawa, Akira Tokumura","Kaori Ueda, Masanori Yoshihara, Michiyasu Nakao, Tamotsu Tanaka, Shigeki Sano, Kenji Fukuzawa, Akira Tokumura","null","Overproduction of lysophosphatidic acid (LPA) by lysophospholipase D/autotaxin (lysoPLD/ATX) is postulated to be involved in the promotion of cancer and atherosclerosis. A lysoPLD inhibitor may be utilized to ameliorate the LPA-related pathological conditions. In this study, a new assay was devised to quantify p-nitrophenol from hydrolysis of chromogenic substrate by serum lysoPLD without tedious lipid extraction procedures. Flavonols, phenolic acids, free fatty acids, and N-acyltyrosines inhibited lysoPLD activity in a micromolar range. They were classified into competitive, noncompetitive, or mixed type inhibitors. The results show that the low hydrophobicity of an inhibitor is a critical factor in its preference for the binding to a noncatalytic binding site over a catalytic binding site. Considering its reported bioavailability and the low dependency of its inhibitory activity on serum dilution, flavonol is likely to be a more effective lysoPLD inhibitor in human blood circulation in vivo than the other inhibitors including LPA.","Overproduction of lysophosphatidic acid (LPA) by lysophospholipase D/autotaxin (lysoPLD/ATX) is postulated to be involved in the promotion of cancer and atherosclerosis. A lysoPLD inhibitor may be utilized to ameliorate the LPA-related pathological conditions. In this study, a new assay was devised to quantify p-nitrophenol from hydrolysis of chromogenic substrate by serum lysoPLD without tedious lipid extraction procedures. Flavonols, phenolic acids, free fatty acids, and N-acyltyrosines inhibited lysoPLD activity in a micromolar range. They were classified into competitive, noncompetitive, or mixed type inhibitors. The results show that the low hydrophobicity of an inhibitor is a critical factor in its preference for the binding to a noncatalytic binding site over a catalytic binding site. Considering its reported bioavailability and the low dependency of its inhibitory activity on serum dilution, flavonol is likely to be a more effective lysoPLD inhibitor in human blood circulation in vivo than the other inhibitors including LPA.","null","null","2010-05-26","Journal of Agricultural and Food Chemistry","Journal of Agricultural and Food Chemistry","Vol.58","No.10","6053","6063","eng","true","true","scientific_journal","null","null","10.1021/jf904155a","1520-5118","null","null","null","null","null"
"A clean-up technology for the simultaneous determination of lysophosphatidic acid and sphingosine-1-phosphate by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using a phosphate-capture molecule, Phos-tag.","A clean-up technology for the simultaneous determination of lysophosphatidic acid and sphingosine-1-phosphate by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using a phosphate-capture molecule, Phos-tag.","Jun-ichi Morishige, Mai Urikura, Haruko Takagi, Kaoru Hirano, Tohru Koike, Tamotsu Tanaka, Kiyoshi Satouchi","Jun-ichi Morishige, Mai Urikura, Haruko Takagi, Kaoru Hirano, Tohru Koike, Tamotsu Tanaka, Kiyoshi Satouchi","null","Lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are growth factor-like lipids having a phosphate group. The concentrations of these mediator lipids in blood are considered to be potential biomarkers for early detection of cancer or vascular diseases. Here, we report a method for simultaneous determination of LPA and S1P using Phos-tag, a zinc complex that specifically binds to a phosphate-monoester group. Although both LPA and S1P are hydrophilic compounds, we found that they acquire hydrophobic properties when they form complexes with Phos-tag. Based on this finding, we developed a method for the enrichment of LPA and S1P from biological samples. The first partition in a two-phase solvent system consisting of chloroform/methanol/water (1:1:0.9, v/v/v) is conducted for the removal of lipids. LPA and S1P are specifically extracted as Phos-tag complexes at the second partition by adding Phos-tag. The Phos-tag complexes of LPA and S1P are detectable by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and quantifiable based on the relative intensities of ions using 17:0 LPA and C17 S1P as internal standards. The protocol was validated by analyses of these mediator lipids in calf serum, a rat brain and a lung. The clean-up protocol is rapid, requires neither thin-layer chromatography (TLC) nor liquid chromatography (LC), and is applicable to both blood and solid tissue samples. We believe that our protocol will be useful for a routine analysis of LPA and S1P in many clinical samples.","Lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are growth factor-like lipids having a phosphate group. The concentrations of these mediator lipids in blood are considered to be potential biomarkers for early detection of cancer or vascular diseases. Here, we report a method for simultaneous determination of LPA and S1P using Phos-tag, a zinc complex that specifically binds to a phosphate-monoester group. Although both LPA and S1P are hydrophilic compounds, we found that they acquire hydrophobic properties when they form complexes with Phos-tag. Based on this finding, we developed a method for the enrichment of LPA and S1P from biological samples. The first partition in a two-phase solvent system consisting of chloroform/methanol/water (1:1:0.9, v/v/v) is conducted for the removal of lipids. LPA and S1P are specifically extracted as Phos-tag complexes at the second partition by adding Phos-tag. The Phos-tag complexes of LPA and S1P are detectable by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and quantifiable based on the relative intensities of ions using 17:0 LPA and C17 S1P as internal standards. The protocol was validated by analyses of these mediator lipids in calf serum, a rat brain and a lung. The clean-up protocol is rapid, requires neither thin-layer chromatography (TLC) nor liquid chromatography (LC), and is applicable to both blood and solid tissue samples. We believe that our protocol will be useful for a routine analysis of LPA and S1P in many clinical samples.","null","null","2010-04-15","Rapid Communications in Mass Spectrometry: RCM","Rapid Communications in Mass Spectrometry: RCM","Vol.24","No.7","1075","1084","eng","true","true","scientific_journal","null","null","10.1002/rcm.4484","1097-0231","null","null","null","null","null"
"S1P3-mediated cardiac fibrosis in sphingosine kinase 1 transgenic mice involves reactive oxygen species.","S1P3-mediated cardiac fibrosis in sphingosine kinase 1 transgenic mice involves reactive oxygen species.","Noriko Takuwa, Sei-Ichiro Ohkura, Shin-Ichiro Takashima, Keisuke Ohtani, Yasuo Okamoto, Tamotsu Tanaka, Kaoru Hirano, Soichiro Usui, Fei Wang, Wa Du, Kazuaki Yoshioka, Yoshiko Banno, Motoko Sasaki, Ikuyo Ichi, Miwa Okamura, Naotoshi Sugimoto, Kiyomi Mizugishi, Yasuni Nakanuma, Isao Ishii, Masayuki Takamura, Shuichi Kaneko, Shosuke Kojo, Kiyoshi Satouchi, Kunitoshi Mitumori, Jerold Chun, Yoh Takuwa","Noriko Takuwa, Sei-Ichiro Ohkura, Shin-Ichiro Takashima, Keisuke Ohtani, Yasuo Okamoto, Tamotsu Tanaka, Kaoru Hirano, Soichiro Usui, Fei Wang, Wa Du, Kazuaki Yoshioka, Yoshiko Banno, Motoko Sasaki, Ikuyo Ichi, Miwa Okamura, Naotoshi Sugimoto, Kiyomi Mizugishi, Yasuni Nakanuma, Isao Ishii, Masayuki Takamura, Shuichi Kaneko, Shosuke Kojo, Kiyoshi Satouchi, Kunitoshi Mitumori, Jerold Chun, Yoh Takuwa","null","AIMS: Sphingosine kinase 1 (SPHK1), its product sphingosine-1-phosphate (S1P), and S1P receptor subtypes have been suggested to play protective roles for cardiomyocytes in animal models of ischaemic preconditioning and cardiac ischaemia/reperfusion injury. To get more insight into roles for SPHK1 in vivo, we have generated SPHK1-transgenic (TG) mice and analysed the cardiac phenotype. METHODS AND RESULTS: SPHK1-TG mice overexpressed SPHK1 in diverse tissues, with a nearly 20-fold increase in enzymatic activity. The TG mice grew normally with normal blood chemistry, cell counts, heart rate, and blood pressure. Unexpectedly, TG mice with high but not low expression levels of SPHK1 developed progressive myocardial degeneration and fibrosis, with upregulation of embryonic genes, elevated RhoA and Rac1 activity, stimulation of Smad3 phosphorylation, and increased levels of oxidative stress markers. Treatment of juvenile TG mice with pitavastatin, an established inhibitor of the Rho family G proteins, or deletion of S1P3, a major myocardial S1P receptor subtype that couples to Rho GTPases and transactivates Smad signalling, both inhibited cardiac fibrosis with concomitant inhibition of SPHK1-dependent Smad-3 phosphorylation. In addition, the anti-oxidant N-2-mercaptopropyonylglycine, which reduces reactive oxygen species (ROS), also inhibited cardiac fibrosis. In in vivo ischaemia/reperfusion injury, the size of myocardial infarct was 30% decreased in SPHK1-TG mice compared with wild-type mice. CONCLUSION: These results suggest that chronic activation of SPHK1-S1P signalling results in both pathological cardiac remodelling through ROS mediated by S1P3 and favourable cardioprotective effects.","AIMS: Sphingosine kinase 1 (SPHK1), its product sphingosine-1-phosphate (S1P), and S1P receptor subtypes have been suggested to play protective roles for cardiomyocytes in animal models of ischaemic preconditioning and cardiac ischaemia/reperfusion injury. To get more insight into roles for SPHK1 in vivo, we have generated SPHK1-transgenic (TG) mice and analysed the cardiac phenotype. METHODS AND RESULTS: SPHK1-TG mice overexpressed SPHK1 in diverse tissues, with a nearly 20-fold increase in enzymatic activity. The TG mice grew normally with normal blood chemistry, cell counts, heart rate, and blood pressure. Unexpectedly, TG mice with high but not low expression levels of SPHK1 developed progressive myocardial degeneration and fibrosis, with upregulation of embryonic genes, elevated RhoA and Rac1 activity, stimulation of Smad3 phosphorylation, and increased levels of oxidative stress markers. Treatment of juvenile TG mice with pitavastatin, an established inhibitor of the Rho family G proteins, or deletion of S1P3, a major myocardial S1P receptor subtype that couples to Rho GTPases and transactivates Smad signalling, both inhibited cardiac fibrosis with concomitant inhibition of SPHK1-dependent Smad-3 phosphorylation. In addition, the anti-oxidant N-2-mercaptopropyonylglycine, which reduces reactive oxygen species (ROS), also inhibited cardiac fibrosis. In in vivo ischaemia/reperfusion injury, the size of myocardial infarct was 30% decreased in SPHK1-TG mice compared with wild-type mice. CONCLUSION: These results suggest that chronic activation of SPHK1-S1P signalling results in both pathological cardiac remodelling through ROS mediated by S1P3 and favourable cardioprotective effects.","null","null","2009-09-15","Cardiovascular Research","Cardiovascular Research","Vol.85","No.3","484","493","eng","true","true","scientific_journal","null","null","10.1093/cvr/cvp312","1755-3245","null","null","null","null","null"
"Formation of lysophosphatidic acid, a wound-healing lipid, during digestion of cabbage leaves.","Formation of lysophosphatidic acid, a wound-healing lipid, during digestion of cabbage leaves.","Tamotsu Tanaka, Gou Horiuchi, Megumi Matsuoka, Kaoru Hirano, Akira Tokumura, Tohru Koike, Kiyoshi Satouchi","Tamotsu Tanaka, Gou Horiuchi, Megumi Matsuoka, Kaoru Hirano, Akira Tokumura, Tohru Koike, Kiyoshi Satouchi","null","Lysophosphatidic acid (LPA) is a lipid mediator that plays a role in the process of wound healing in animal tissues, including the digestive tract. We determined LPA in several foodstuffs, and found that cabbage leaves were the richest source of LPA. We also found that, at 22 and 195 nmol/g (wet weight), LPA and phosphatidic acid (PA) were respectively formed during mastication of raw cabbage leaves and that the resulting PA was converted to LPA by pancreatic phospholipase A(2). The lipid extract obtained from ground cabbage leaves promoted the proliferation of Swiss 3T3 fibroblasts and the motility of HGC-27 cells, stomach-derived epithelial-like cells, at physiologically relevant concentrations. These activities of cabbage lipids were inhibited by Ki16425, an LPA-receptor antagonist. LPA formed during the digestion of cabbage leaves may be one of the components in the beneficial effect of ingested cabbage on a damaged digestive tract.","Lysophosphatidic acid (LPA) is a lipid mediator that plays a role in the process of wound healing in animal tissues, including the digestive tract. We determined LPA in several foodstuffs, and found that cabbage leaves were the richest source of LPA. We also found that, at 22 and 195 nmol/g (wet weight), LPA and phosphatidic acid (PA) were respectively formed during mastication of raw cabbage leaves and that the resulting PA was converted to LPA by pancreatic phospholipase A(2). The lipid extract obtained from ground cabbage leaves promoted the proliferation of Swiss 3T3 fibroblasts and the motility of HGC-27 cells, stomach-derived epithelial-like cells, at physiologically relevant concentrations. These activities of cabbage lipids were inhibited by Ki16425, an LPA-receptor antagonist. LPA formed during the digestion of cabbage leaves may be one of the components in the beneficial effect of ingested cabbage on a damaged digestive tract.","null","null","2009-06-07","Bioscience, Biotechnology, and Biochemistry","Bioscience, Biotechnology, and Biochemistry","Vol.73","No.6","1293","1300","eng","true","true","scientific_journal","null","null","10.1271/bbb.80813","1347-6947","null","http://ci.nii.ac.jp/naid/10027542114/","null","null","null"
"Inhibitory effect of juniperonic acid (Delta-5c,11c,14c,17c-20:4, omega-3) on bombesin-induced proliferation of Swiss 3T3 cells.","Inhibitory effect of juniperonic acid (Delta-5c,11c,14c,17c-20:4, omega-3) on bombesin-induced proliferation of Swiss 3T3 cells.","Jun-ichi Morishige, Naoki Amano, Kaoru Hirano, Hiroaki Nishio, Tamotsu Tanaka, Kiyoshi Satouchi","Jun-ichi Morishige, Naoki Amano, Kaoru Hirano, Hiroaki Nishio, Tamotsu Tanaka, Kiyoshi Satouchi","null","Juniperonic acid (Delta-5c,11c,14c,17c-20:4, JA) is a polymethylene-interrupted (PMI) fatty acid that occurs in Biota orientalis. In this study, we found that JA has an antiproliferative activity. Swiss 3T3 cells were preloaded with fatty acids before stimulation with bombesin, a mitogenic neuropeptide, and proliferation of the cells was assessed by [(3)H]thymidine incorporation. Preloading of linoleic acid (Delta-9c,12c-18:2) significantly enhanced bombesin-induced proliferation. In contrast, preloading of eicosapentaenoic acid (Delta-5c,8c,11c,14c,17c-20:5, EPA) suppressed proliferation. Likewise, cells preloaded with JA showed a significantly curtailed response to bombesin. The antiproliferative potency of JA was equivalent to that of EPA. Sciadonic acid (Delta-5c,11c,14c-20:3), an omega-6 analogue of JA did not show antiproliferative activity, suggesting the importance of the omega-3 double bond rather than the PMI structure. The EPA-like activity of JA may be involved in the pharmaceutical activity of biota seeds, a psychoactive Chinese traditional medicine.","Juniperonic acid (Delta-5c,11c,14c,17c-20:4, JA) is a polymethylene-interrupted (PMI) fatty acid that occurs in Biota orientalis. In this study, we found that JA has an antiproliferative activity. Swiss 3T3 cells were preloaded with fatty acids before stimulation with bombesin, a mitogenic neuropeptide, and proliferation of the cells was assessed by [(3)H]thymidine incorporation. Preloading of linoleic acid (Delta-9c,12c-18:2) significantly enhanced bombesin-induced proliferation. In contrast, preloading of eicosapentaenoic acid (Delta-5c,8c,11c,14c,17c-20:5, EPA) suppressed proliferation. Likewise, cells preloaded with JA showed a significantly curtailed response to bombesin. The antiproliferative potency of JA was equivalent to that of EPA. Sciadonic acid (Delta-5c,11c,14c-20:3), an omega-6 analogue of JA did not show antiproliferative activity, suggesting the importance of the omega-3 double bond rather than the PMI structure. The EPA-like activity of JA may be involved in the pharmaceutical activity of biota seeds, a psychoactive Chinese traditional medicine.","null","null","2008-09","Biological & Pharmaceutical Bulletin","Biological & Pharmaceutical Bulletin","Vol.31","No.9","1786","1789","eng","true","true","scientific_journal","null","null","10.1248/bpb.31.1786","0918-6158","null","null","null","null","null"
"クローブ(syzygium aromaticum L.)熱水抽出物の高脂肪食摂餌マウス脂質代謝に及ぼす影響","Effect of Hot Water Extracts from Clove (Syzygium aromaticum L.) on Lipid Metabolism in High-Fat Diet Fed Mice","吉積 一真, 平野 薫, 富 裕孝, 田中 保, 里内 清","吉積 一真, 平野 薫, 富 裕孝, Tamotsu Tanaka, 里内 清","null","The effect of a hot water extract of clove (Syzygium aromaticum L.) on serum and hepatic lipids, and fecal triglyceride levels was investigated in mice. The addition of 1.0% (w/w) of the clove hot water extract to a high-fat diet containing 60% (w/w) lard for 28 days significantly increased fecal triglyceride levels, significantly reduced plasma insulin and hepatic total cholesterol levels, and tended to reduce plasma and hepatic triglyceride levels compared with mice given a high-fat diet without the clove hot water extract. Semiquantitative real time reverse transcription-polymerase chain reaction analysis demonstrated that addition of the clove hot water extract to a high-fat diet increased gene expression of medium-chain acyl-CoA dehydrogenase and reduced gene expression of fatty acid synthase in the liver. In addition, the clove hot water extract dose-dependently inhibited lipase activity in vitro. It is therefore suggested that the improvement action of lipid metabolism in mice fed a high-fat diet originated from the activation of lipid oxidation, the inactivation of fatty acid synthesis in the liver, and the inhibition of lipolysis in the small intestine.","The effect of a hot water extract of clove (Syzygium aromaticum L.) on serum and hepatic lipids, and fecal triglyceride levels was investigated in mice. The addition of 1.0% (w/w) of the clove hot water extract to a high-fat diet containing 60% (w/w) lard for 28 days significantly increased fecal triglyceride levels, significantly reduced plasma insulin and hepatic total cholesterol levels, and tended to reduce plasma and hepatic triglyceride levels compared with mice given a high-fat diet without the clove hot water extract. Semiquantitative real time reverse transcription-polymerase chain reaction analysis demonstrated that addition of the clove hot water extract to a high-fat diet increased gene expression of medium-chain acyl-CoA dehydrogenase and reduced gene expression of fatty acid synthase in the liver. In addition, the clove hot water extract dose-dependently inhibited lipase activity in vitro. It is therefore suggested that the improvement action of lipid metabolism in mice fed a high-fat diet originated from the activation of lipid oxidation, the inactivation of fatty acid synthesis in the liver, and the inhibition of lipolysis in the small intestine.","null","null","2007-08-20","生薬学雑誌","The Japanese Journal of Pharmacognosy","Vol.61","No.2","79","85","jpn","true","null","scientific_journal","null","null","null","1349-9114","null","http://ci.nii.ac.jp/naid/110008729790/","null","null","null"
"Metabolic pathway that produces essential fatty acids from polymethylene-interrupted polyunsaturated fatty acids in animal cells","Metabolic pathway that produces essential fatty acids from polymethylene-interrupted polyunsaturated fatty acids in animal cells","Tamotsu Tanaka, J. Morishige, D. Iwawaki, T. Fukuhara, K. Satouchi","Tamotsu Tanaka, J. Morishige, D. Iwawaki, T. Fukuhara, K. Satouchi","null","Sciadonic acid (20:3 Delta-5,11,14) and juniperonic acid (20:4 Delta-5,11,14,17) are polyunsaturated fatty acids (PUFAs) that lack the Delta-8 double bond of arachidonic acid (20:4 Delta-5,8,11,14) and eicosapentaenoic acid (20:5 Delta-5,8,11,14,17), respectively. Here, we demonstrate that these conifer oil-derived PUFAs are metabolized to essential fatty acids in animal cells. When Swiss 3T3 cells were cultured with sciadonic acid, linoleic acid (18:2 Delta-9,12) accumulated in the cells to an extent dependent on the concentration of sciadonic acid. At the same time, a small amount of 16:2 Delta-7,10 appeared in the cellular lipids. Both 16:2 Delta-7,10 and linoleic acid accumulated in sciadonic acid-supplemented CHO cells, but not in peroxisome-deficient CHO cells. We confirmed that 16:2 Delta-7,10 was effectively elongated to linoleic acid in rat liver microsomes. These results indicate that sciadonic acid was partially degraded to 16:2 Delta-7,10 by two cycles of beta-oxidation in peroxisomes, then elongated to linoleic acid in microsomes. Supplementation of Swiss 3T3 cells with juniperonic acid, an n-3 analogue of sciadonic acid, induced accumulation of alpha-linolenic acid (18:3 Delta-9,12,15) in cellular lipids, suggesting that juniperonic acid was metabolized in a similar manner to sciadonic acid. This PUFA remodeling is thought to be a process that converts unsuitable fatty acids into essential fatty acids required by animals.","Sciadonic acid (20:3 Delta-5,11,14) and juniperonic acid (20:4 Delta-5,11,14,17) are polyunsaturated fatty acids (PUFAs) that lack the Delta-8 double bond of arachidonic acid (20:4 Delta-5,8,11,14) and eicosapentaenoic acid (20:5 Delta-5,8,11,14,17), respectively. Here, we demonstrate that these conifer oil-derived PUFAs are metabolized to essential fatty acids in animal cells. When Swiss 3T3 cells were cultured with sciadonic acid, linoleic acid (18:2 Delta-9,12) accumulated in the cells to an extent dependent on the concentration of sciadonic acid. At the same time, a small amount of 16:2 Delta-7,10 appeared in the cellular lipids. Both 16:2 Delta-7,10 and linoleic acid accumulated in sciadonic acid-supplemented CHO cells, but not in peroxisome-deficient CHO cells. We confirmed that 16:2 Delta-7,10 was effectively elongated to linoleic acid in rat liver microsomes. These results indicate that sciadonic acid was partially degraded to 16:2 Delta-7,10 by two cycles of beta-oxidation in peroxisomes, then elongated to linoleic acid in microsomes. Supplementation of Swiss 3T3 cells with juniperonic acid, an n-3 analogue of sciadonic acid, induced accumulation of alpha-linolenic acid (18:3 Delta-9,12,15) in cellular lipids, suggesting that juniperonic acid was metabolized in a similar manner to sciadonic acid. This PUFA remodeling is thought to be a process that converts unsuitable fatty acids into essential fatty acids required by animals.","null","null","2007-04-20","The FEBS Journal","The FEBS Journal","Vol.274","No.11","2728","2737","eng","true","null","scientific_journal","null","null","10.1111/j.1742-4658.2007.05807.x","1742-464X","null","null","null","null","null"
"CGI-58 facilitates lipolysis on lipid droplets but is not involved in the vesiculation of lipid droplets caused by hormonal stimulation","CGI-58 facilitates lipolysis on lipid droplets but is not involved in the vesiculation of lipid droplets caused by hormonal stimulation","T. Yamaguchi, N. Omatsu, E. Morimoto, H. Nakashima, Tamotsu Tanaka","T. Yamaguchi, N. Omatsu, E. Morimoto, H. Nakashima, Tamotsu Tanaka","null","A lipid droplet (LD)-associated protein, perilipin, is a critical regulator of lipolysis in adipocytes. We previously showed that Comparative Gene Identification-58 (CGI-58), a product of the causal gene of Chanarin-Dorfman syndrome, interacts with perilipin on LDs. In this study, we investigated the function of CGI-58 using RNA interference. Notably, CGI-58 knockdown caused an abnormal accumulation of LDs in both 3T3-L1 preadipocytes and Hepa1 hepatoma cells. CGI-58 knockdown did not influence the differentiation of 3T3-L1 adipocytes but reduced the activity of both basal and cAMP-dependent protein kinase-stimulated lipolysis. In vitro studies showed that CGI-58 itself does not have lipase/esterase activity, but it enhanced the activity of adipose triglyceride lipase. Upon lipolytic stimulation, endogenous CGI-58 was rapidly dispersed from LDs into the cytosol along with small particulate structures. This shift in localization depends on the phosphorylation of perilipin, because phosphorylated perilipin lost the ability to bind CGI-58. During lipolytic activation, LDs in adipocytes vesiculate into micro-LDs. Using coherent anti-Stokes Raman scattering microscopy, we pursued the formation of micro-LDs in single cells, which seemed to occur in cytoplasmic regions distant from the large central LDs. CGI-58 is not required for this process. Thus, CGI-58 facilitates lipolysis in cooperation with perilipin and other factors, including lipases.","A lipid droplet (LD)-associated protein, perilipin, is a critical regulator of lipolysis in adipocytes. We previously showed that Comparative Gene Identification-58 (CGI-58), a product of the causal gene of Chanarin-Dorfman syndrome, interacts with perilipin on LDs. In this study, we investigated the function of CGI-58 using RNA interference. Notably, CGI-58 knockdown caused an abnormal accumulation of LDs in both 3T3-L1 preadipocytes and Hepa1 hepatoma cells. CGI-58 knockdown did not influence the differentiation of 3T3-L1 adipocytes but reduced the activity of both basal and cAMP-dependent protein kinase-stimulated lipolysis. In vitro studies showed that CGI-58 itself does not have lipase/esterase activity, but it enhanced the activity of adipose triglyceride lipase. Upon lipolytic stimulation, endogenous CGI-58 was rapidly dispersed from LDs into the cytosol along with small particulate structures. This shift in localization depends on the phosphorylation of perilipin, because phosphorylated perilipin lost the ability to bind CGI-58. During lipolytic activation, LDs in adipocytes vesiculate into micro-LDs. Using coherent anti-Stokes Raman scattering microscopy, we pursued the formation of micro-LDs in single cells, which seemed to occur in cytoplasmic regions distant from the large central LDs. CGI-58 is not required for this process. Thus, CGI-58 facilitates lipolysis in cooperation with perilipin and other factors, including lipases.","null","null","2007-02-17","Journal of Lipid Research","Journal of Lipid Research","Vol.48","No.5","1078","1089","eng","true","null","scientific_journal","null","null","10.1194/jlr.M600493-JLR200","0022-2275","null","null","null","null","null"
"Production of bioactive lysophosphatidic acid by lysophospholipase D in hen egg white","Production of bioactive lysophosphatidic acid by lysophospholipase D in hen egg white","Junichi Morishige, Kanako Touchika, Tamotsu Tanaka, Kiyoshi Satouchi, Kenji Fukuzawa, Akira Tokumura","Junichi Morishige, Kanako Touchika, Tamotsu Tanaka, Kiyoshi Satouchi, Kenji Fukuzawa, Akira Tokumura","null","Lysophosphatidic acid (LPA), a lysophospholipid mediator, is produced extracellularly by lysophospholipase D (lysoPLD) secreted in several animal body fluids including blood plasma. Previously, we reported that hen egg white contains polyunsaturated fatty acid-rich LPA. In this study, we examined whether lysoPLD is involved in the production of LPA in hen egg white. LysoPLD activity was measured by determining LPA and choline by mass spectrometric and enzyme-linked fluorometric analyses, respectively. LysoPLD increased with increased dilution of egg white, indicating that one or more components of egg white strongly inhibit its lysoPLD activity. This dilution-dependent increase in the lysoPLD activity was masked by co-incubation of the egg white with lysozyme, a major protein in hen egg white. Furthermore, addition of Zn(2+), Mn(2+), Ni(2+), or Co(2+) to diluted egg white altered preference patterns of lysoPLD toward choline-containing substrates. In particular, the egg white lysoPLD activity was greatly increased when Co(2+) was added. The cation-requirement of lysoPLD activity in hen egg white resembled that of plasma autotaxin (ATX)/lysoPLD. Western blot analysis revealed that egg white contained a protein that was immunostained with anti-ATX antibody. These results suggested that LPA in hen egg white is produced from lysophospholipids, especially LPC, by the action of ATX/lysoPLD, possibly originating from hen oviduct fluid.","Lysophosphatidic acid (LPA), a lysophospholipid mediator, is produced extracellularly by lysophospholipase D (lysoPLD) secreted in several animal body fluids including blood plasma. Previously, we reported that hen egg white contains polyunsaturated fatty acid-rich LPA. In this study, we examined whether lysoPLD is involved in the production of LPA in hen egg white. LysoPLD activity was measured by determining LPA and choline by mass spectrometric and enzyme-linked fluorometric analyses, respectively. LysoPLD increased with increased dilution of egg white, indicating that one or more components of egg white strongly inhibit its lysoPLD activity. This dilution-dependent increase in the lysoPLD activity was masked by co-incubation of the egg white with lysozyme, a major protein in hen egg white. Furthermore, addition of Zn(2+), Mn(2+), Ni(2+), or Co(2+) to diluted egg white altered preference patterns of lysoPLD toward choline-containing substrates. In particular, the egg white lysoPLD activity was greatly increased when Co(2+) was added. The cation-requirement of lysoPLD activity in hen egg white resembled that of plasma autotaxin (ATX)/lysoPLD. Western blot analysis revealed that egg white contained a protein that was immunostained with anti-ATX antibody. These results suggested that LPA in hen egg white is produced from lysophospholipids, especially LPC, by the action of ATX/lysoPLD, possibly originating from hen oviduct fluid.","null","null","2007-01-19","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1771","No.4","491","499","eng","true","null","scientific_journal","null","null","10.1016/j.bbalip.2007.01.005","1388-1981","null","null","null","null","null"
"Lupane-type saponins from leaves of Acanthopanax sessiliflorus and their inhibitory activity on pancreatic lipase","Lupane-type saponins from leaves of Acanthopanax sessiliflorus and their inhibitory activity on pancreatic lipase","Kazuma Yoshizumi, Kaoru Hirano, Hidehiro Ando, Tamotsu Tanaka, Junji Terao","Kazuma Yoshizumi, Kaoru Hirano, Hidehiro Ando, Tamotsu Tanaka, Junji Terao","null","Three known saponins, chiisanoside, 11-deoxyisochiisanoside, and isochiisanoside, and one novel saponin, 3,4-seco-4(23),20(29)-lupadiene-3,28-dioic acid 28-O-alpha-l-rhamnopyranosyl (1-->4)-beta-d-glucopyranosyl (1-->6)-beta-d-glucopyranoside, referred to as sessiloside, were isolated from a hot water extract of Acanthopanax sessiliflorus leaves. All of these saponins were lupane-type triterpene triglycosides, and their concentrations were 4.1, 1.0, 0.5, and 0.4% (w/w) of the total extract, respectively. Sessiloside and chiisanoside inhibited pancreatic lipase activity in vitro, and addition of the saponin-rich fraction to a high-fat diet suppressed the body weight gain of mice. The possibility of application of the lupane-type saponins from A. sessiliflorus leaves to the treatment of obesity is discussed.","Three known saponins, chiisanoside, 11-deoxyisochiisanoside, and isochiisanoside, and one novel saponin, 3,4-seco-4(23),20(29)-lupadiene-3,28-dioic acid 28-O-alpha-l-rhamnopyranosyl (1-->4)-beta-d-glucopyranosyl (1-->6)-beta-d-glucopyranoside, referred to as sessiloside, were isolated from a hot water extract of Acanthopanax sessiliflorus leaves. All of these saponins were lupane-type triterpene triglycosides, and their concentrations were 4.1, 1.0, 0.5, and 0.4% (w/w) of the total extract, respectively. Sessiloside and chiisanoside inhibited pancreatic lipase activity in vitro, and addition of the saponin-rich fraction to a high-fat diet suppressed the body weight gain of mice. The possibility of application of the lupane-type saponins from A. sessiliflorus leaves to the treatment of obesity is discussed.","null","null","2006-01-25","Journal of Agricultural and Food Chemistry","Journal of Agricultural and Food Chemistry","Vol.54","No.2","335","341","eng","true","null","scientific_journal","null","null","10.1021/jf052047f","0021-8561","null","null","null","null","null"
"Production and protein kinase C activation of diacylglycerols with polymethylene-interrupted PUFA residues","Production and protein kinase C activation of diacylglycerols with polymethylene-interrupted PUFA residues","J. Morishige, Y. Takai, K. Hirano, Tamotsu Tanaka, K. Satouchi","J. Morishige, Y. Takai, K. Hirano, Tamotsu Tanaka, K. Satouchi","null","Sciadonic acid (20:3, delta-5c,11 c,14c) is a polymethylene-interrupted PUFA (PMI-PUFA) that is present in conifer seeds and known to be incorporated into animal cells and to accumulate in membrane PI as a substitute for arachidonate. In this study, we investigated whether PI having sciadonate could serve as source of DAG that could activate protein kinase C (PKC). When Swiss 3T3 cells cultured with sciadonic acid were stimulated with 100 nM of bombesin, 1-stearoyl-2-sciadonoyl-glycerol (G) and 1-stearoyl-2-arachidonoyl-G were produced. The net increments of these two molecular species of DAG reflected the levels of the two molecular species in the PI in the cells. When cells cultured with juniperonic acid (20:4, delta-5c,11c,14c,17c) were stimulated, 1-stearoyl-2-juniperonoyl-G was produced in proportion to the level of this molecular species in PI in the cells. We also examined PKC activation by synthetic DAG using a partially purified PKC fraction from rat brain and found that both 1-stearoyl-2-sciadonoyl-G and 1-stearoyl-2-juniperonoyl-G could activate PKC comparably to 1 -stearoyl-2-arachidonoyl-G. These results indicate that 1-stearoyl-PI having these C20 PMI-PUFA residues can serve as sources of potential signaling molecules.","Sciadonic acid (20:3, delta-5c,11 c,14c) is a polymethylene-interrupted PUFA (PMI-PUFA) that is present in conifer seeds and known to be incorporated into animal cells and to accumulate in membrane PI as a substitute for arachidonate. In this study, we investigated whether PI having sciadonate could serve as source of DAG that could activate protein kinase C (PKC). When Swiss 3T3 cells cultured with sciadonic acid were stimulated with 100 nM of bombesin, 1-stearoyl-2-sciadonoyl-glycerol (G) and 1-stearoyl-2-arachidonoyl-G were produced. The net increments of these two molecular species of DAG reflected the levels of the two molecular species in the PI in the cells. When cells cultured with juniperonic acid (20:4, delta-5c,11c,14c,17c) were stimulated, 1-stearoyl-2-juniperonoyl-G was produced in proportion to the level of this molecular species in PI in the cells. We also examined PKC activation by synthetic DAG using a partially purified PKC fraction from rat brain and found that both 1-stearoyl-2-sciadonoyl-G and 1-stearoyl-2-juniperonoyl-G could activate PKC comparably to 1 -stearoyl-2-arachidonoyl-G. These results indicate that 1-stearoyl-PI having these C20 PMI-PUFA residues can serve as sources of potential signaling molecules.","null","null","2005-02","Lipids","Lipids","Vol.40","No.2","155","162","eng","true","null","scientific_journal","null","null","10.1007/s11745-005-1370-8","0024-4201","null","null","null","null","null"
"Quantitative analysis of lysophosphatidic acid by time-of-flight mass spectrometry using a phosphate-capture molecule","Quantitative analysis of lysophosphatidic acid by time-of-flight mass spectrometry using a phosphate-capture molecule","Tamotsu Tanaka, Hideki Tsutsui, Kaoru Hirano, Tohru Koike, Akira Tokumura, Kiyoshi Satouchi","Tamotsu Tanaka, Hideki Tsutsui, Kaoru Hirano, Tohru Koike, Akira Tokumura, Kiyoshi Satouchi","null","Lysophosphatidic acid (LPA) is a lipid mediator that may play an important role in wound healing, embryonic development, and progression of cancer. Here, we report a procedure for the quantification of LPA by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The method is based on a characteristic mass shift with total charge change (from -2 to +1) of the phosphate species due to 1:1 complexation of LPA(2-) with a dinuclear zinc (II) complex [1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-olato dizinc(II) complex; Zn(2)L(3+)] at physiological pH. The monocationic complex [LPA(2-)-Zn(2)L(3+)](+) was detected in the positive mode, in which no other signal of cation adducts of LPA(2-) was observed. The detection limit of 18:1 LPA by this method was 0.1 pmol on a sample plate. The intensity ratio of [LPA(2-)-Zn(2)L(3+)](+) against an internal standard [17:0 LPA(2-)-Zn(2)L(3+)](+) increased linearly with their molar ratio. Based on the relative intensities of complex ions, we determined the amounts of LPA homologs in an egg white by this method; the results obtained were in good agreement with those by gas liquid chromatography. This sensitive and convenient procedure for LPA-specific detection is useful for the quantification of LPA homologs occurring in biological materials.","Lysophosphatidic acid (LPA) is a lipid mediator that may play an important role in wound healing, embryonic development, and progression of cancer. Here, we report a procedure for the quantification of LPA by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The method is based on a characteristic mass shift with total charge change (from -2 to +1) of the phosphate species due to 1:1 complexation of LPA(2-) with a dinuclear zinc (II) complex [1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-olato dizinc(II) complex; Zn(2)L(3+)] at physiological pH. The monocationic complex [LPA(2-)-Zn(2)L(3+)](+) was detected in the positive mode, in which no other signal of cation adducts of LPA(2-) was observed. The detection limit of 18:1 LPA by this method was 0.1 pmol on a sample plate. The intensity ratio of [LPA(2-)-Zn(2)L(3+)](+) against an internal standard [17:0 LPA(2-)-Zn(2)L(3+)](+) increased linearly with their molar ratio. Based on the relative intensities of complex ions, we determined the amounts of LPA homologs in an egg white by this method; the results obtained were in good agreement with those by gas liquid chromatography. This sensitive and convenient procedure for LPA-specific detection is useful for the quantification of LPA homologs occurring in biological materials.","null","null","2004-11","Journal of Lipid Research","Journal of Lipid Research","Vol.45","No.11","2145","2150","eng","true","null","scientific_journal","null","null","10.1194/jlr.D400010-JLR200","0022-2275","null","http://www.jlr.org/cgi/content/full/45/11/2145","null","null","null"
"Production of 1,2-didocosahexaenoyl phosphatidylcholine by bonito muscle lysophosphatidylcholine/transacylase","Production of 1,2-didocosahexaenoyl phosphatidylcholine by bonito muscle lysophosphatidylcholine/transacylase","K. Hirano, H. Matsui, Tamotsu Tanaka, F. Matsuura, K. Satouchi","K. Hirano, H. Matsui, Tamotsu Tanaka, F. Matsuura, K. Satouchi","null","1,2-Didocosahexaenoyl phosphatidylcholine (PC), which has highly unsaturated fatty acid at both sn-1 and sn-2 positions of glycerol, is a characteristic molecular species of bonito muscle. To examine the involvement of a de novo route in its synthesis, the molecular species of phosphatidic acid (PA) were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using a 1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-olato dizinc(II) complex, a novel phosphate-capture molecule. However, 1,2-didocosahexaenoyl species could not be detected. Next, 1,2-didocosahexaenoyl PC synthesis by the cytosolic lysophosphatidylcholine (LPC)/transacylase was examined using endogenous LPC from bonito muscle, in which the 2-docosahexaenoyl species is abundant. The LPC/transacylase synthesized 1,2-didocosahexaenoyl PC as the most abundant molecular species. For further characterization, the LPC/transacylase was purified to homogeneity from the 100,000 x g supernatant of bonito muscle. The isolated LPC/transacylase is a labile glycoprotein with molecular mass of 52 kDa including a 5-kDa sugar moiety. The LPC/transacylase showed a PC synthesis (transacylase activity) below and above the critical micelle concentration of substrate LPC, and fatty acid release (lysophospholipase activity) was always smaller than the transacylase activity, even with a monomeric substrate. These results suggest that the LPC/transacylase is responsible for the synthesis of 1,2-didocosahexaenoyl PC.","1,2-Didocosahexaenoyl phosphatidylcholine (PC), which has highly unsaturated fatty acid at both sn-1 and sn-2 positions of glycerol, is a characteristic molecular species of bonito muscle. To examine the involvement of a de novo route in its synthesis, the molecular species of phosphatidic acid (PA) were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using a 1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-olato dizinc(II) complex, a novel phosphate-capture molecule. However, 1,2-didocosahexaenoyl species could not be detected. Next, 1,2-didocosahexaenoyl PC synthesis by the cytosolic lysophosphatidylcholine (LPC)/transacylase was examined using endogenous LPC from bonito muscle, in which the 2-docosahexaenoyl species is abundant. The LPC/transacylase synthesized 1,2-didocosahexaenoyl PC as the most abundant molecular species. For further characterization, the LPC/transacylase was purified to homogeneity from the 100,000 x g supernatant of bonito muscle. The isolated LPC/transacylase is a labile glycoprotein with molecular mass of 52 kDa including a 5-kDa sugar moiety. The LPC/transacylase showed a PC synthesis (transacylase activity) below and above the critical micelle concentration of substrate LPC, and fatty acid release (lysophospholipase activity) was always smaller than the transacylase activity, even with a monomeric substrate. These results suggest that the LPC/transacylase is responsible for the synthesis of 1,2-didocosahexaenoyl PC.","null","null","2004-10","The Journal of Biochemistry","The Journal of Biochemistry","Vol.136","No.4","477","483","eng","true","null","scientific_journal","null","null","10.1093/jb/mvh145","0021-924X","null","null","null","null","null"
"Mechanisms of accumulation of arachidonate in phosphatidylinositol in yellowtail: A comparative study of acylation systems of phospholipids in rat","Mechanisms of accumulation of arachidonate in phosphatidylinositol in yellowtail: A comparative study of acylation systems of phospholipids in rat","Tamotsu Tanaka, D. Iwawaki, M. Sakamoto, Y. Takai, K. Satouchi","Tamotsu Tanaka, D. Iwawaki, M. Sakamoto, Y. Takai, K. Satouchi","null","It is known that phosphatidylinositol (PtdIns) contains abundant arachidonate and is composed mainly of 1-stearoyl-2-arachidonoyl species in mammals. We investigated if this characteristic of PtdIns applies to the PtdIns from yellowtail (Seriola quinqueradiata), a marine fish. In common with phosphatidylcholine (PtdCho), phosphatidylethanolamine (PtdEtn) and phosphatidylserine (PtdSer) from brain, heart, liver, spleen, kidney and ovary, the predominant polyunsaturated fatty acid was docosahexaenoic acid, and levels of arachidonic acid were less than 4.5% (PtdCho), 7.5% (PtdEtn) and 3.0% (PtdSer) in these tissues. In striking contrast, arachidonic acid made up 17.6%, 31.8%, 27.8%, 26.1%, 25.4% and 33.5% of the fatty acid composition of PtdIns from brain, heart, liver, spleen, kidney and ovary, respectively. The most abundant molecular species of PtdIns in all these tissues was 1-stearoyl-2-arachidonoyl. Assay of acyltransferase in liver microsomes of yellowtail showed that arachidonic acid was incorporated into PtdIns more effectively than docosahexaenoic acid and that the latter inhibited incorporation of arachidonic acid into PtdCho without inhibiting the utilization of arachidonic acid for PtdIns. This effect of docosahexaenoic acid was not observed in similar experiments using rat liver microsomes and is thought to contribute to the exclusive utilization of arachidonic acid for acylation to PtdIns in yellowtail. Inositolphospholipids and their hydrolysates are known to act as signaling molecules in cells. The conserved hydrophobic structure of PtdIns (the 1-stearoyl-2-arachidonoyl moiety) may have physiological significance not only in mammals but also in fish.","It is known that phosphatidylinositol (PtdIns) contains abundant arachidonate and is composed mainly of 1-stearoyl-2-arachidonoyl species in mammals. We investigated if this characteristic of PtdIns applies to the PtdIns from yellowtail (Seriola quinqueradiata), a marine fish. In common with phosphatidylcholine (PtdCho), phosphatidylethanolamine (PtdEtn) and phosphatidylserine (PtdSer) from brain, heart, liver, spleen, kidney and ovary, the predominant polyunsaturated fatty acid was docosahexaenoic acid, and levels of arachidonic acid were less than 4.5% (PtdCho), 7.5% (PtdEtn) and 3.0% (PtdSer) in these tissues. In striking contrast, arachidonic acid made up 17.6%, 31.8%, 27.8%, 26.1%, 25.4% and 33.5% of the fatty acid composition of PtdIns from brain, heart, liver, spleen, kidney and ovary, respectively. The most abundant molecular species of PtdIns in all these tissues was 1-stearoyl-2-arachidonoyl. Assay of acyltransferase in liver microsomes of yellowtail showed that arachidonic acid was incorporated into PtdIns more effectively than docosahexaenoic acid and that the latter inhibited incorporation of arachidonic acid into PtdCho without inhibiting the utilization of arachidonic acid for PtdIns. This effect of docosahexaenoic acid was not observed in similar experiments using rat liver microsomes and is thought to contribute to the exclusive utilization of arachidonic acid for acylation to PtdIns in yellowtail. Inositolphospholipids and their hydrolysates are known to act as signaling molecules in cells. The conserved hydrophobic structure of PtdIns (the 1-stearoyl-2-arachidonoyl moiety) may have physiological significance not only in mammals but also in fish.","null","null","2003-04","European Journal of Biochemistry","European Journal of Biochemistry","Vol.270","No.7","1466","1473","eng","true","null","scientific_journal","null","null","10.1046/j.1432-1033.2003.03512.x","0014-2956","null","null","null","null","null"
"A lipase-inhibiting protein from lipoxygenase-deficient soybean seeds","A lipase-inhibiting protein from lipoxygenase-deficient soybean seeds","K. Satouchi, Y. Kodama, K. Hirano, Tamotsu Tanaka, H. Iwamoto","K. Satouchi, Y. Kodama, K. Hirano, Tamotsu Tanaka, H. Iwamoto","null","A lipase-inhibiting protein was isolated from lipoxygenase (LOX)-deficient soybean seeds. The molecular mass of the protein was 56.0-kDa and the N-terminal amino acid was blocked. The protein was identified by peptide mass fingerprinting in combination with matrix-assisted laser desorption ionization/time-of-flight mass spectrometry. The masses of the lysyl endopeptidase-digested peptides of the 56.0-kDa inhibiting protein were almost identical to the calculated masses of the theoretically predicted lysyl endopeptidase-treated peptides of beta-amylase from soybean seed. In a previous paper (Biosci. Biotechnol. Biochem., 62, 1498-1503, 1998), we reported that LOX-1, an isozyme of soybean seed LOX, inhibited hydrolysis of soybean oil by pancreatic lipase. Purified beta-amylase also inhibited lipase activity, although the magnitude of inhibition was weaker than that by LOX-1. Thus, there are at least two lipase-inhibiting proteins, one is a LOX and the other is a beta-amylase, in soybean seed.","A lipase-inhibiting protein was isolated from lipoxygenase (LOX)-deficient soybean seeds. The molecular mass of the protein was 56.0-kDa and the N-terminal amino acid was blocked. The protein was identified by peptide mass fingerprinting in combination with matrix-assisted laser desorption ionization/time-of-flight mass spectrometry. The masses of the lysyl endopeptidase-digested peptides of the 56.0-kDa inhibiting protein were almost identical to the calculated masses of the theoretically predicted lysyl endopeptidase-treated peptides of beta-amylase from soybean seed. In a previous paper (Biosci. Biotechnol. Biochem., 62, 1498-1503, 1998), we reported that LOX-1, an isozyme of soybean seed LOX, inhibited hydrolysis of soybean oil by pancreatic lipase. Purified beta-amylase also inhibited lipase activity, although the magnitude of inhibition was weaker than that by LOX-1. Thus, there are at least two lipase-inhibiting proteins, one is a LOX and the other is a beta-amylase, in soybean seed.","null","null","2002-10","Bioscience, Biotechnology, and Biochemistry","Bioscience, Biotechnology, and Biochemistry","Vol.66","No.10","2154","2160","eng","true","null","scientific_journal","null","null","10.1271/bbb.66.2154","0916-8451","null","null","null","null","null"
"Human platelets respond differentially to lysophosphatidic acids having a highly unsaturated fatty acyl group and alkyl ether-linked lysophosphatidic acids","Human platelets respond differentially to lysophosphatidic acids having a highly unsaturated fatty acyl group and alkyl ether-linked lysophosphatidic acids","Akira Tokumura, Junya Shinomiya, Seishi Kishimoto, Tamotsu Tanaka, Kentaro Kogure, Takayuki Sugiura, Kiyoshi Satouchi, Keizo Waku, Kenji Fukuzawa","Akira Tokumura, Junya Shinomiya, Seishi Kishimoto, Tamotsu Tanaka, Kentaro Kogure, Takayuki Sugiura, Kiyoshi Satouchi, Keizo Waku, Kenji Fukuzawa","null","Lysophosphatidic acid (LPA) is a physiological agonist that is produced by lysophospholipase D, phospholipase A(1) and phospholipase A(2) in the blood of animals. It exerts diverse biological actions on a broad range of animal cells. Specific receptors for this important agonist have been characterized. In this investigation, for the first time we prepared LPAs having a highly unsaturated fatty acyl group, such as the eicosapentaenoyl or docosahexaenoyl residue, and their acetylated derivatives. Human platelets aggregated more potently in response to the highly unsaturated acyl-LPAs than to LPAs with a C(18) fatty acyl group, such as an oleoyl group, while alkyl ether-linked LPAs (alkyl-LPA) had much stronger aggregating activity. Two positional isomers of LPAs with an arachidonoyl, eicosapentaenoyl or docosahexaenoyl group had equipotent aggregatory activity as well as the positional isomers of their acetylated analogues, indicating that putative LPA receptors could not distinguish the difference between the positional isomers. We found that platelet preparations from two individuals showed no aggregatory response to alkyl-LPAs, although they contained mRNAs for known LPA receptors in the following order of expression level: endothelial differentiation gene (Edg)-4>Edg-7>Edg-2. We also obtained evidence that 2-(p-amylcinnamoyl)amino-4-chlorobenzoic acid (ONO-RS-082), a phospholipase A(2) inhibitor, potentiated alkyl-LPA-induced platelet aggregation, but inhibited highly unsaturated acyl-LPA-induced platelet aggregation. These results indicated that human platelets express acyl-LPA-selective and alkyl-LPA-selective receptors on their plasma membrane.","Lysophosphatidic acid (LPA) is a physiological agonist that is produced by lysophospholipase D, phospholipase A(1) and phospholipase A(2) in the blood of animals. It exerts diverse biological actions on a broad range of animal cells. Specific receptors for this important agonist have been characterized. In this investigation, for the first time we prepared LPAs having a highly unsaturated fatty acyl group, such as the eicosapentaenoyl or docosahexaenoyl residue, and their acetylated derivatives. Human platelets aggregated more potently in response to the highly unsaturated acyl-LPAs than to LPAs with a C(18) fatty acyl group, such as an oleoyl group, while alkyl ether-linked LPAs (alkyl-LPA) had much stronger aggregating activity. Two positional isomers of LPAs with an arachidonoyl, eicosapentaenoyl or docosahexaenoyl group had equipotent aggregatory activity as well as the positional isomers of their acetylated analogues, indicating that putative LPA receptors could not distinguish the difference between the positional isomers. We found that platelet preparations from two individuals showed no aggregatory response to alkyl-LPAs, although they contained mRNAs for known LPA receptors in the following order of expression level: endothelial differentiation gene (Edg)-4>Edg-7>Edg-2. We also obtained evidence that 2-(p-amylcinnamoyl)amino-4-chlorobenzoic acid (ONO-RS-082), a phospholipase A(2) inhibitor, potentiated alkyl-LPA-induced platelet aggregation, but inhibited highly unsaturated acyl-LPA-induced platelet aggregation. These results indicated that human platelets express acyl-LPA-selective and alkyl-LPA-selective receptors on their plasma membrane.","null","null","2002-08-01","The Biochemical Journal","The Biochemical Journal","Vol.365","No.Pt 3","617","628","eng","true","null","scientific_journal","null","null","10.1042/BJ20020348","0264-6021","null","null","null","null","null"
"Metabolic characterization of sciadonic acid (5c,11c,14c-eicosatrienoic acid) as an effective substitute for arachidonate of phosphatidylinositol","Metabolic characterization of sciadonic acid (5c,11c,14c-eicosatrienoic acid) as an effective substitute for arachidonate of phosphatidylinositol","Tamotsu Tanaka, J. Morishige, T. Takimoto, Y. Takai, K. Satouchi","Tamotsu Tanaka, J. Morishige, T. Takimoto, Y. Takai, K. Satouchi","null","Sciadonic acid (20:3 Delta-5,11,14) is an n-6 series trienoic acid that lacks the Delta8 double bond of arachidonic acid. This fatty acid is not converted to arachidonic acid in higher animals. In this study, we characterized the metabolic behavior of sciadonic acid in the process of acylation to phospholipid of HepG2 cells. One of the characteristics of fatty acid compositions of phospholipids in sciadonic acid-supplemented cells is a higher proportion of sciadonic acid in phosphatidylinositol (PtdIns) (27.4%) than in phosphatidylethanolamine (PtdEtn) (23.2%), phosphatidylcholine (PtdCho) (17.3%) and phosphatidylserine (PtdSer) (20.1%). Similarly, the proportion of arachidonic acid was higher in PtdIns (35.8%) than in PtdEtn (29.1%), PtdSer (18.2%) and PtdCho (20.2%) in arachidonic-acid-supplemented cells. The extensive accumulation of sciadonic acid in PtdIns resulted in the enrichment of newly formed 1-stearoyl-2-sciadonoyl molecular species (38%) in PtdIns and caused the reduction in the level of pre-existing arachidonic-acid-containing molecular species. The kinetics of incorporation of sciadonic acid to PtdEtn, PtdSer and PtdIns of cells were similar to those of arachidonic acid. In contrast to sciadonic acid, neither eicosapentaenoic acid (20:5 Delta-5,8,11,14,17) nor juniperonic acid (20:4 Delta-5,11,14,17) accumulated in the PtdIns fraction. Rather, these n-3 series polyunsaturated fatty acids, once incorporated into PtdIns, tended to be excluded from PtdIns. In addition, the level of arachidonic-acid-containing PtdIns molecular species remained unchanged by eicosapentaenoic-acid-supplementation. These results suggest that sciadonic acid or sciadonic-acid-containing glycerides are metabolized in a similar manner to arachidonic acid or arachidonic-acid-containing glyceride in the biosynthesis of PtdIns and that sciadonic acid can effectively modify the molecular species composition of PtdIns in HepG2 cells. In this regard, sciadonic acid will be an interesting experimental tool to clarify the significance of arachidonic acid-residue of PtdIns-origin bioactive lipids.","Sciadonic acid (20:3 Delta-5,11,14) is an n-6 series trienoic acid that lacks the Delta8 double bond of arachidonic acid. This fatty acid is not converted to arachidonic acid in higher animals. In this study, we characterized the metabolic behavior of sciadonic acid in the process of acylation to phospholipid of HepG2 cells. One of the characteristics of fatty acid compositions of phospholipids in sciadonic acid-supplemented cells is a higher proportion of sciadonic acid in phosphatidylinositol (PtdIns) (27.4%) than in phosphatidylethanolamine (PtdEtn) (23.2%), phosphatidylcholine (PtdCho) (17.3%) and phosphatidylserine (PtdSer) (20.1%). Similarly, the proportion of arachidonic acid was higher in PtdIns (35.8%) than in PtdEtn (29.1%), PtdSer (18.2%) and PtdCho (20.2%) in arachidonic-acid-supplemented cells. The extensive accumulation of sciadonic acid in PtdIns resulted in the enrichment of newly formed 1-stearoyl-2-sciadonoyl molecular species (38%) in PtdIns and caused the reduction in the level of pre-existing arachidonic-acid-containing molecular species. The kinetics of incorporation of sciadonic acid to PtdEtn, PtdSer and PtdIns of cells were similar to those of arachidonic acid. In contrast to sciadonic acid, neither eicosapentaenoic acid (20:5 Delta-5,8,11,14,17) nor juniperonic acid (20:4 Delta-5,11,14,17) accumulated in the PtdIns fraction. Rather, these n-3 series polyunsaturated fatty acids, once incorporated into PtdIns, tended to be excluded from PtdIns. In addition, the level of arachidonic-acid-containing PtdIns molecular species remained unchanged by eicosapentaenoic-acid-supplementation. These results suggest that sciadonic acid or sciadonic-acid-containing glycerides are metabolized in a similar manner to arachidonic acid or arachidonic-acid-containing glyceride in the biosynthesis of PtdIns and that sciadonic acid can effectively modify the molecular species composition of PtdIns in HepG2 cells. In this regard, sciadonic acid will be an interesting experimental tool to clarify the significance of arachidonic acid-residue of PtdIns-origin bioactive lipids.","null","null","2001-09","European Journal of Biochemistry","European Journal of Biochemistry","Vol.268","No.18","4928","4939","eng","true","null","scientific_journal","null","null","10.1046/j.0014-2956.2001.02423.x","0014-2956","null","null","null","null","null"
"Occurrence of a novel cannabimimetic molecule 2-sciadonoylglycerol (2-eicosa-5',11',14'-trienoylglycerol) in the umbrella pine Sciadopitys veticillata seeds","Occurrence of a novel cannabimimetic molecule 2-sciadonoylglycerol (2-eicosa-5',11',14'-trienoylglycerol) in the umbrella pine Sciadopitys veticillata seeds","S. Nakane, Tamotsu Tanaka, K. Satouchi, Y. Kobayashi, T. Sugiura","S. Nakane, Tamotsu Tanaka, K. Satouchi, Y. Kobayashi, T. Sugiura","null","The umbrella pine Sciadopitys verticillata seeds were found to contain a substantial amount (16.7 nmol/g) of sciadonic acid (all-cis-5,11,14-eicosatrienoic acid)-containing 2-monoacylglycerol, i.e., 2-sciadonoylglycerol (2-eicosa-5',11',14'-trienoylglycerol). Because the structure of 2-sciadonoylglycerol closely resembles that of 2-arachidonoylglycerol, the endogenous natural ligand for the cannabinoid receptor, we examined whether or not 2-sciadonoylglycerol exhibits cannabimimetic activity using NG108-15 neuroblastomaxglioma hybrid cells which express the cannabinoid CB1 receptor. We found that 2-sciadonoylglycerol induces rapid transient elevation of intracellular free Ca2+ concentration in NG108-15 cells through a cannabinoid CBI receptor-dependent mechanism similar to the case of 2-arachidonoylglycerol, yet the activity of 2-sciadonoylglycerol was apparently lower than that of 2-arachidonoylglycerol. The activity of 2-sciadonoylglycerol was detectable from 3-10 nM, reaching a maximum at around 10 microM. To our knowledge, this is the first report showing the occurrence of a cannabimimetic monoacylglycerol in higher plants.","The umbrella pine Sciadopitys verticillata seeds were found to contain a substantial amount (16.7 nmol/g) of sciadonic acid (all-cis-5,11,14-eicosatrienoic acid)-containing 2-monoacylglycerol, i.e., 2-sciadonoylglycerol (2-eicosa-5',11',14'-trienoylglycerol). Because the structure of 2-sciadonoylglycerol closely resembles that of 2-arachidonoylglycerol, the endogenous natural ligand for the cannabinoid receptor, we examined whether or not 2-sciadonoylglycerol exhibits cannabimimetic activity using NG108-15 neuroblastomaxglioma hybrid cells which express the cannabinoid CB1 receptor. We found that 2-sciadonoylglycerol induces rapid transient elevation of intracellular free Ca2+ concentration in NG108-15 cells through a cannabinoid CBI receptor-dependent mechanism similar to the case of 2-arachidonoylglycerol, yet the activity of 2-sciadonoylglycerol was apparently lower than that of 2-arachidonoylglycerol. The activity of 2-sciadonoylglycerol was detectable from 3-10 nM, reaching a maximum at around 10 microM. To our knowledge, this is the first report showing the occurrence of a cannabimimetic monoacylglycerol in higher plants.","null","null","2000-06","Biological & Pharmaceutical Bulletin","Biological & Pharmaceutical Bulletin","Vol.23","No.6","758","761","eng","true","null","scientific_journal","null","null","null","0918-6158","null","null","null","null","null"
"Purification and regiospecificity of multiple enzyme activities of phospholipase A1 from bonito muscle","Purification and regiospecificity of multiple enzyme activities of phospholipase A1 from bonito muscle","K. Hirano, E. Okada, Tamotsu Tanaka, K. Satouchi","K. Hirano, E. Okada, Tamotsu Tanaka, K. Satouchi","null","Phospholipase A(1) (PLA(1)), which catalyzes the hydrolysis of the sn-1 ester bond of diacyl phospholipids, was purified from 100,000 x g supernatant of bonito muscle to homogeneity by ammonium-sulfate precipitation and four consecutive column chromatographies (DEAE anion-exchange, ether-Toyopeal, hydroxylapatite and Toyopeal HW 50S columns). The final preparation showed a single band above the 67-kDa molecular marker on SDS-PAGE, and the molecular mass was determined to be 71.5 kDa by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using bovine serum albumin as a standard for calibration. The N-terminal 8 amino residues were determined to be Ala-Pro-Ala-Glu-Lys-Val-Lys-Try. Regiospecificity of multiple enzyme activities of the PLA(1) was examined using positionally defined synthetic phosphatidylcholine (PC) and lysophosphatidylcholines (LPC). An acyl ester bond at the sn-1 position of PC was exclusively hydrolyzed by phospholipase activity, and 1-acyl LPC was cleaved to fatty acid and glycerophosphocholine by lysophospholipase (LPL) activity. However, the positional isomer, 2-acyl LPC was a poor substrate for LPL activity. PC/transacylation activity was also observed when excess 2-acyl LPC was supplied in the reaction mixture, and fatty acid at the sn-1 position of donor PC was transferred to the sn-1 position of acceptor LPC. These results demonstrate that the multiple enzyme activities of PLA(1), this is lysophospholipase, transacylase as well as phospholipase, have a strict regiospecificity at the sn-1 position of substrates.","Phospholipase A(1) (PLA(1)), which catalyzes the hydrolysis of the sn-1 ester bond of diacyl phospholipids, was purified from 100,000 x g supernatant of bonito muscle to homogeneity by ammonium-sulfate precipitation and four consecutive column chromatographies (DEAE anion-exchange, ether-Toyopeal, hydroxylapatite and Toyopeal HW 50S columns). The final preparation showed a single band above the 67-kDa molecular marker on SDS-PAGE, and the molecular mass was determined to be 71.5 kDa by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using bovine serum albumin as a standard for calibration. The N-terminal 8 amino residues were determined to be Ala-Pro-Ala-Glu-Lys-Val-Lys-Try. Regiospecificity of multiple enzyme activities of the PLA(1) was examined using positionally defined synthetic phosphatidylcholine (PC) and lysophosphatidylcholines (LPC). An acyl ester bond at the sn-1 position of PC was exclusively hydrolyzed by phospholipase activity, and 1-acyl LPC was cleaved to fatty acid and glycerophosphocholine by lysophospholipase (LPL) activity. However, the positional isomer, 2-acyl LPC was a poor substrate for LPL activity. PC/transacylation activity was also observed when excess 2-acyl LPC was supplied in the reaction mixture, and fatty acid at the sn-1 position of donor PC was transferred to the sn-1 position of acceptor LPC. These results demonstrate that the multiple enzyme activities of PLA(1), this is lysophospholipase, transacylase as well as phospholipase, have a strict regiospecificity at the sn-1 position of substrates.","null","null","2000-01","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids","Vol.1483","No.3","325","333","eng","true","null","scientific_journal","null","null","10.1016/S1388-1981(99)00190-0","1388-1981","null","null","null","null","null"
"Non-methylene-interrupted polyunsaturated fatty acids: Effective substitute for arachidonate of phosphatidylinositol","Non-methylene-interrupted polyunsaturated fatty acids: Effective substitute for arachidonate of phosphatidylinositol","Tamotsu Tanaka, S. Izuwa, K. Tanaka, D. Yamamoto, K. Satouchi","Tamotsu Tanaka, S. Izuwa, K. Tanaka, D. Yamamoto, K. Satouchi","null","In mammalian tissues and cells, a characteristic of phosphatidylinositol (PI) is a high abundance of arachidonic acid (AA) relative to the other phospholipids. In this study, we investigated the effects of supplementation of several polyunsaturated fatty acids (PUFAs) on the AA concentration of the PI fraction using a cultured cell system. Neither alpha-linolenic acid nor eicosapentaenoic acid supplement reduced the level of AA in PI of HepG2 cells. In contrast to the n-3 series PUFAs, adding podocarpic acid (20:3, Delta-5,11,14) and pinolenic acid (18:3, Delta-5,9,12) reduced the AA content of the PI fraction from a control value of 15.9% to 7.0 and 8.7%, respectively. In the experiments with pinolenic acid, selective and significant accumulation of 20:3 (Delta-7,11,14), the chain-elongated metabolite of pinolenic acid, was observed in the PI fraction. On the other hand, adding columbinic acid (18:3, Delta-5t,9,12) had no effect on AA content of the PI fraction. Because both podocarpic acid and pinolenic acid are non-methylene-interrupted fatty acids (NMIFAs) that are not converted to AA metabolically, these NMIFAs may be interesting experimental tools for research on the function of PI-origin bioactive lipids.","In mammalian tissues and cells, a characteristic of phosphatidylinositol (PI) is a high abundance of arachidonic acid (AA) relative to the other phospholipids. In this study, we investigated the effects of supplementation of several polyunsaturated fatty acids (PUFAs) on the AA concentration of the PI fraction using a cultured cell system. Neither alpha-linolenic acid nor eicosapentaenoic acid supplement reduced the level of AA in PI of HepG2 cells. In contrast to the n-3 series PUFAs, adding podocarpic acid (20:3, Delta-5,11,14) and pinolenic acid (18:3, Delta-5,9,12) reduced the AA content of the PI fraction from a control value of 15.9% to 7.0 and 8.7%, respectively. In the experiments with pinolenic acid, selective and significant accumulation of 20:3 (Delta-7,11,14), the chain-elongated metabolite of pinolenic acid, was observed in the PI fraction. On the other hand, adding columbinic acid (18:3, Delta-5t,9,12) had no effect on AA content of the PI fraction. Because both podocarpic acid and pinolenic acid are non-methylene-interrupted fatty acids (NMIFAs) that are not converted to AA metabolically, these NMIFAs may be interesting experimental tools for research on the function of PI-origin bioactive lipids.","null","null","1999-11-02","Biochemical and Biophysical Research Communications","Biochemical and Biophysical Research Communications","Vol.264","No.3","683","688","eng","true","null","scientific_journal","null","null","10.1006/bbrc.1999.1559","0006-291X","null","null","null","null","null"
"Biosynthesis of 1,2-dieicosapentaenoyl-sn-glycero-3-phosphocholine in Caenorhabditis elegans","Biosynthesis of 1,2-dieicosapentaenoyl-sn-glycero-3-phosphocholine in Caenorhabditis elegans","Tamotsu Tanaka, S. Izuwa, K. Tanaka, D. Yamamoto, K. Satouchi","Tamotsu Tanaka, S. Izuwa, K. Tanaka, D. Yamamoto, K. Satouchi","null","Previously, we showed that lowering the growth temperature increased the level of eicosapentaenoic acid (EPA) in the phosphatidylcholine (PtdCho) of Caenorhabditis elegans. In this study, we investigated the molecular species composition of PtdCho of C. elegans, with an emphasis on EPA-containing species. C. elegans contained a substantial amount of 1,2-dipolyunsaturated fatty acid-containing PtdCho (1,2-diPUFA-PtdCho) species, such as arachidonic acid/EPA and EPA/EPA, which are unusual phospholipids in higher animals. The EPA/EPA-PtdCho content was significantly increased in C. elegans grown at a low temperature. To examine the possibility that the acyltransferase activity involved in the remodeling of phospholipids accounts for the production of 1,2-diPUFA-PtdCho, we investigated the substrate specificity of this enzyme in C. elegans and found that it did not exhibit a preference for saturated fatty acid for acylation to the sn-1 position of PtdCho. The efficacy of the esterification of EPA to the sn-1 position was almost equal to that of stearic acid. The lack of preference for a saturated fatty acid for acylation to the sn-1 position of PtdCho is thought to result in the existence of the unusual 1,2-diEPA-PtdCho in C. elegans.","Previously, we showed that lowering the growth temperature increased the level of eicosapentaenoic acid (EPA) in the phosphatidylcholine (PtdCho) of Caenorhabditis elegans. In this study, we investigated the molecular species composition of PtdCho of C. elegans, with an emphasis on EPA-containing species. C. elegans contained a substantial amount of 1,2-dipolyunsaturated fatty acid-containing PtdCho (1,2-diPUFA-PtdCho) species, such as arachidonic acid/EPA and EPA/EPA, which are unusual phospholipids in higher animals. The EPA/EPA-PtdCho content was significantly increased in C. elegans grown at a low temperature. To examine the possibility that the acyltransferase activity involved in the remodeling of phospholipids accounts for the production of 1,2-diPUFA-PtdCho, we investigated the substrate specificity of this enzyme in C. elegans and found that it did not exhibit a preference for saturated fatty acid for acylation to the sn-1 position of PtdCho. The efficacy of the esterification of EPA to the sn-1 position was almost equal to that of stearic acid. The lack of preference for a saturated fatty acid for acylation to the sn-1 position of PtdCho is thought to result in the existence of the unusual 1,2-diEPA-PtdCho in C. elegans.","null","null","1999-07","European Journal of Biochemistry","European Journal of Biochemistry","Vol.263","No.1","189","194","eng","true","null","scientific_journal","null","null","10.1046/j.1432-1327.1999.00480.x","0014-2956","null","http://ci.nii.ac.jp/naid/80011179991/","null","null","null"
"Methylene-interrupted double bond in polyunsaturated fatty acid is essential structure for metabolism by fatty acid chain elongation system of rat liver","Methylene-interrupted double bond in polyunsaturated fatty acid is essential structure for metabolism by fatty acid chain elongation system of rat liver","Tamotsu Tanaka, T. Hattori, M. Kouchi, K. Hirano, K. Satouchi","Tamotsu Tanaka, T. Hattori, M. Kouchi, K. Hirano, K. Satouchi","null","Some plant oils contain non-methylene-interrupted polyunsaturated fatty acids (NMIFAs). Pinolenic acid (all cis delta-5,9,12/18:3) and columbinic acid (trans,cis,cis delta-5,9,12/18:3) are NMIFAs that exist in pine seed oil and columbine seed oil, respectively. We investigated the double bond position of fatty acid recognized by the fatty acid chain elongation system (FACES) of rat liver using NMIFAs as experimental tools. In the total elongation assay, amounts of C2 unit chain-elongated metabolites of pinolenic acid and columbinic acid were 32% and 11%, respectively, compared to that of gamma-linolenic (all cis delta-6,9,12/18:3) as the substrate. In the condensation reaction assay, the rate limiting step of FACES, the conversion rates of pinolenic acid and columbinic acid to the corresponding C20 beta-keto fatty acids were 19% and 9% of that of gamma-linolenic acid, respectively. The formation of elongated metabolite of podocarpic acid (all cis delta-5,11,14/20:3) was only 7% of that of arachidonic acid (all cis delta-5,8,11,14/20:4). From these results it was concluded that the condensing enzyme of FACES could recognize the methylene-interrupted cis double bond structure vicinal to the carboxyl group in the fatty acid molecule.","Some plant oils contain non-methylene-interrupted polyunsaturated fatty acids (NMIFAs). Pinolenic acid (all cis delta-5,9,12/18:3) and columbinic acid (trans,cis,cis delta-5,9,12/18:3) are NMIFAs that exist in pine seed oil and columbine seed oil, respectively. We investigated the double bond position of fatty acid recognized by the fatty acid chain elongation system (FACES) of rat liver using NMIFAs as experimental tools. In the total elongation assay, amounts of C2 unit chain-elongated metabolites of pinolenic acid and columbinic acid were 32% and 11%, respectively, compared to that of gamma-linolenic (all cis delta-6,9,12/18:3) as the substrate. In the condensation reaction assay, the rate limiting step of FACES, the conversion rates of pinolenic acid and columbinic acid to the corresponding C20 beta-keto fatty acids were 19% and 9% of that of gamma-linolenic acid, respectively. The formation of elongated metabolite of podocarpic acid (all cis delta-5,11,14/20:3) was only 7% of that of arachidonic acid (all cis delta-5,8,11,14/20:4). From these results it was concluded that the condensing enzyme of FACES could recognize the methylene-interrupted cis double bond structure vicinal to the carboxyl group in the fatty acid molecule.","null","null","1998-08-28","Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism","Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism","Vol.1393","No.2-3","299","306","eng","true","null","scientific_journal","null","null","10.1016/S0005-2760(98)00084-8","0005-2760","null","null","null","null","null"
"Cytosolic lysophosphatidylcholine/transacylase in the production of dipolyunsaturated phosphatidylcholine in bonito muscle","Cytosolic lysophosphatidylcholine/transacylase in the production of dipolyunsaturated phosphatidylcholine in bonito muscle","K. Hirano, H. Ito, H. Morihara, Tamotsu Tanaka, K. Satouchi","K. Hirano, H. Ito, H. Morihara, Tamotsu Tanaka, K. Satouchi","null","null","null","null","null","1998","FEBS Lett.","FEBS Lett.","Vol.437","null","193","196","eng","true","null","scientific_journal","null","null","null","null","null","null","null","null","null"
"Lipoxygenase-1 from soybean seed inhibiting the activity of pancreatic lipase","Lipoxygenase-1 from soybean seed inhibiting the activity of pancreatic lipase","K. Satouchi, K. Hirano, O. Fijino, M. Ikoma, Tamotsu Tanaka","K. Satouchi, K. Hirano, O. Fijino, M. Ikoma, Tamotsu Tanaka","null","null","null","null","null","1998","Biosci. Biotech. Biochem.","Biosci. Biotech. Biochem.","Vol.62","null","1498","1503","eng","true","null","scientific_journal","null","null","null","null","null","null","null","null","null"
"Properties of phospholipase A1/transacylase in the white muscle of Bonito Euthynnus pelamis (Linnaeus)","Properties of phospholipase A1/transacylase in the white muscle of Bonito Euthynnus pelamis (Linnaeus)","K. Hirano, A. Tanaka, K. Yoshizumi, Tamotsu Tanaka, K. Satouchi","K. Hirano, A. Tanaka, K. Yoshizumi, Tamotsu Tanaka, K. Satouchi","null","The properties of phospholipase A1 (PLA1) obtained from the white muscle of bonito, Euthynnus pelamis (Linnaeus), were examined. The PLA1 activity had a pH optimum from 6.5 to 7.0 for phosphatidylcholine (PC), and calcium ion was not required. The optimum temperature was from 20 to 30 degrees C. When a fatty alcohol was used as an acceptor, a wax ester was produced by transferring a fatty acid at the sn-1 position of the donor's PC. The maximum production of lysophosphatidylcholine was shifted by 0.5 pH units to the acidic side and the pH optimum of wax ester synthesis was from 6.0 to 6.5. The synthesis was independent of calcium ion and Coenzyme A. The transacylation was also observed when 1-lyso-2-acyl-sn-glycero-3-phosphocholine was used as an acceptor. Fatty acid at the sn-1 position of the donor PC was transferred to the unoccupied hydroxy group of the acceptor at the sn-1 position. When 2,3-dipalmitoyl-sn-glycero-1-phosphocholine was used as the acyl donor, a similar amount of palmitic acid was transferred as in the case of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine. However, 1-acyl-2-lyso-sn -glycero-3-phosphocholine, a positional isomer, was a poor acceptor. These results indicate that the transacylation by the PLA1 from bonito muscle is not stereospecific, but is position-specific both for the acyl donor and acceptor.","The properties of phospholipase A1 (PLA1) obtained from the white muscle of bonito, Euthynnus pelamis (Linnaeus), were examined. The PLA1 activity had a pH optimum from 6.5 to 7.0 for phosphatidylcholine (PC), and calcium ion was not required. The optimum temperature was from 20 to 30 degrees C. When a fatty alcohol was used as an acceptor, a wax ester was produced by transferring a fatty acid at the sn-1 position of the donor's PC. The maximum production of lysophosphatidylcholine was shifted by 0.5 pH units to the acidic side and the pH optimum of wax ester synthesis was from 6.0 to 6.5. The synthesis was independent of calcium ion and Coenzyme A. The transacylation was also observed when 1-lyso-2-acyl-sn-glycero-3-phosphocholine was used as an acceptor. Fatty acid at the sn-1 position of the donor PC was transferred to the unoccupied hydroxy group of the acceptor at the sn-1 position. When 2,3-dipalmitoyl-sn-glycero-1-phosphocholine was used as the acyl donor, a similar amount of palmitic acid was transferred as in the case of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine. However, 1-acyl-2-lyso-sn -glycero-3-phosphocholine, a positional isomer, was a poor acceptor. These results indicate that the transacylation by the PLA1 from bonito muscle is not stereospecific, but is position-specific both for the acyl donor and acceptor.","null","null","1997-12","The Journal of Biochemistry","The Journal of Biochemistry","Vol.122","No.6","1160","1166","eng","true","null","scientific_journal","null","null","null","0021-924X","null","null","null","null","null"
"Purification and gas chromatographic-mass spectrometric characterization of non-methylene interrupted fatty acid incorporated in rat liver","Purification and gas chromatographic-mass spectrometric characterization of non-methylene interrupted fatty acid incorporated in rat liver","Tamotsu Tanaka, K. Shibata, H. Hino, T. Murashita, K. Satouchi","Tamotsu Tanaka, K. Shibata, H. Hino, T. Murashita, K. Satouchi","null","A C20 non-methylene interrupted trienoic acid detected in the liver of rat fed with a pine (Pinus koraiensis) seed oil diet was purified by two-step argentation thin-layer chromatography (AgTLC) and characterized by gas chromatography-mass spectrometry (GC-MS). First, a C20 methyl trienoate fraction was obtained from fatty acid methyl esters prepared from rat liver by 5% AgTLC developed with petroleum ether-diethyl ether-acetic acid (70:20:2, v/v) as a solvent system. The fraction was then subjected to AgTLC developed with benzene-acetone-diethyl ether-acetic acid (65:15:15:5, v/v) which could separate non-methylene interrupted fatty acids (NMIFA) from usual MIFAs. The purified C20 NMIFA was partially hydrogenated, and the resulting three kinds of the C20 monoenoate were analyzed by GC-MS after conversion to their dimethyl disulfide (DMDS) adducts. The results revealed that the original C20 non-methylene interrupted trienoic acid detected in the liver of rats fed with a pine seed oil diet was delta-5,11,14/20:3, a minor component of pine seed oil.","A C20 non-methylene interrupted trienoic acid detected in the liver of rat fed with a pine (Pinus koraiensis) seed oil diet was purified by two-step argentation thin-layer chromatography (AgTLC) and characterized by gas chromatography-mass spectrometry (GC-MS). First, a C20 methyl trienoate fraction was obtained from fatty acid methyl esters prepared from rat liver by 5% AgTLC developed with petroleum ether-diethyl ether-acetic acid (70:20:2, v/v) as a solvent system. The fraction was then subjected to AgTLC developed with benzene-acetone-diethyl ether-acetic acid (65:15:15:5, v/v) which could separate non-methylene interrupted fatty acids (NMIFA) from usual MIFAs. The purified C20 NMIFA was partially hydrogenated, and the resulting three kinds of the C20 monoenoate were analyzed by GC-MS after conversion to their dimethyl disulfide (DMDS) adducts. The results revealed that the original C20 non-methylene interrupted trienoic acid detected in the liver of rats fed with a pine seed oil diet was delta-5,11,14/20:3, a minor component of pine seed oil.","null","null","1997-10-24","Journal of Chromatography. B, Biomedical Sciences and Applications","Journal of Chromatography. B, Biomedical Sciences and Applications","Vol.700","No.1-2","1","8","eng","true","null","scientific_journal","null","null","null","1387-2273","null","null","null","null","null"
"Effects of growth temperature on the fatty acid composition of the free-living nematode Caenorhabditis elegans","Effects of growth temperature on the fatty acid composition of the free-living nematode Caenorhabditis elegans","Tamotsu Tanaka, K. Ikita, T. Ashida, Y. Motoyama, K. Satouchi","Tamotsu Tanaka, K. Ikita, T. Ashida, Y. Motoyama, K. Satouchi","null","null","null","null","null","1996","Lipids","Lipids","Vol.31","null","1173","1178","eng","true","null","scientific_journal","null","null","null","null","null","null","null","null","null"
"Platelet activating factor (PAF)-like phospholipids formed during peroxidation of phosphatidylcholines from different foodstuffs","Platelet activating factor (PAF)-like phospholipids formed during peroxidation of phosphatidylcholines from different foodstuffs","Tamotsu Tanaka, Akira Tokumura, H. Tsukatani","Tamotsu Tanaka, Akira Tokumura, H. Tsukatani","null","Previously, we reported that induction of peroxidation of synthetic phosphatidylcholines (PCs) containing a polyunsaturated fatty acid by Fe(2+)-EDTA in the presence of ascorbate resulted in the formation of four types of PCs with an sn-2-oxidatively fragmented acyl group, which had platelet-aggregating activity due to interaction with platelet-activating factor (PAF) receptors. These PCs were compounds with a short-chain monocarboxylate, omega-hydroxymonocarboxylate, dicarboxylate, and dicarboxylate semialdehyde residue, respectively. In this study, we investigated the PAF-like lipids formed during peroxidation of PCs from hen egg yolk, salmon roe, sea urchin eggs, and krill in an FeSO4/EDTA/ascorbate system. The platelet-aggregating activities of these oxidized PCs were all inhibited by FR-900452, an antagonist of PAF. The activity of oxidized krill PC, which was equivalent of 89.8 +/- 8.8 pmol 16:0-PAF/mumol of starting PC, was about 5 times those of oxidized PCs from salmon roe and sea urchin eggs, and about 50 times that of oxidized hen egg yolk PC. The PAF-like phospholipids that had different combinations of long-chain alkyl or acyl groups with one of the above four types of short-chain acyl groups were identified by gas chromatography-mass spectrometry. The results indicated that foodstuffs that are rich in 1-O-alkyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine are potential sources of compounds with high PAF-like activity formed by deleterious lipid peroxidation.","Previously, we reported that induction of peroxidation of synthetic phosphatidylcholines (PCs) containing a polyunsaturated fatty acid by Fe(2+)-EDTA in the presence of ascorbate resulted in the formation of four types of PCs with an sn-2-oxidatively fragmented acyl group, which had platelet-aggregating activity due to interaction with platelet-activating factor (PAF) receptors. These PCs were compounds with a short-chain monocarboxylate, omega-hydroxymonocarboxylate, dicarboxylate, and dicarboxylate semialdehyde residue, respectively. In this study, we investigated the PAF-like lipids formed during peroxidation of PCs from hen egg yolk, salmon roe, sea urchin eggs, and krill in an FeSO4/EDTA/ascorbate system. The platelet-aggregating activities of these oxidized PCs were all inhibited by FR-900452, an antagonist of PAF. The activity of oxidized krill PC, which was equivalent of 89.8 +/- 8.8 pmol 16:0-PAF/mumol of starting PC, was about 5 times those of oxidized PCs from salmon roe and sea urchin eggs, and about 50 times that of oxidized hen egg yolk PC. The PAF-like phospholipids that had different combinations of long-chain alkyl or acyl groups with one of the above four types of short-chain acyl groups were identified by gas chromatography-mass spectrometry. The results indicated that foodstuffs that are rich in 1-O-alkyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine are potential sources of compounds with high PAF-like activity formed by deleterious lipid peroxidation.","null","null","1995-08","Bioscience, Biotechnology, and Biochemistry","Bioscience, Biotechnology, and Biochemistry","Vol.59","No.8","1389","1393","eng","true","null","scientific_journal","null","null","10.1271/bbb.59.1389","0916-8451","null","null","null","null","null"
"Platelet-activating factor and its structural analogues in the earthworm Eisenia foetida","Platelet-activating factor and its structural analogues in the earthworm Eisenia foetida","T. Sugiura, A. Yamashita, N. Kudo, Tamotsu Tanaka, Akira Tokumura","T. Sugiura, A. Yamashita, N. Kudo, Tamotsu Tanaka, Akira Tokumura","null","null","null","null","null","1995","Biochim. Biophys. Acta","Biochim. Biophys. Acta","Vol.1258","null","19","26","eng","true","null","scientific_journal","null","null","null","null","null","null","null","null","null"
"Platelet-aggregating effects of platelet-activating factor-like phospholipids formed by oxidation of phosphatidylcholines containing an sn-2 polyunsaturated fatty acyl group","Platelet-aggregating effects of platelet-activating factor-like phospholipids formed by oxidation of phosphatidylcholines containing an sn-2 polyunsaturated fatty acyl group","Tamotsu Tanaka, H. Iimori, H. Tsukatani, Akira Tokumura","Tamotsu Tanaka, H. Iimori, H. Tsukatani, Akira Tokumura","null","null","null","null","null","1994","Biochim. Biophys. Acta","Biochim. Biophys. Acta","Vol.1210","null","202","208","eng","true","null","scientific_journal","null","null","null","null","null","null","null","null","null"
"Lysophosphatidylcholine from white muscle of bonito Euthynnus pelamis (Linnaeus): involvement of phospholipase A1 activity for its production","Lysophosphatidylcholine from white muscle of bonito Euthynnus pelamis (Linnaeus): involvement of phospholipase A1 activity for its production","K. Satouchi, M. Sakaguchi, M. Shirakawa, K. Hirano, Tamotsu Tanaka","K. Satouchi, M. Sakaguchi, M. Shirakawa, K. Hirano, Tamotsu Tanaka","null","null","null","null","null","1994","Biochim. Biophys. Acta","Biochim. Biophys. Acta","Vol.1214","null","303","308","eng","true","null","scientific_journal","null","null","null","null","null","null","null","null","null"
"Formation of platelet-activating factor-like phospholipids by Fe2+/ascorbate/EDTA-induced lipid peroxidation","Formation of platelet-activating factor-like phospholipids by Fe2+/ascorbate/EDTA-induced lipid peroxidation","Tamotsu Tanaka, H. Minamino, S. Unezaki, H. Tsukatani, Akira Tokumura","Tamotsu Tanaka, H. Minamino, S. Unezaki, H. Tsukatani, Akira Tokumura","null","null","null","null","null","1994","Biochim. Biophys. Acta","Biochim. Biophys. Acta","Vol.1210","null","202","208","eng","true","null","scientific_journal","null","null","null","null","null","null","null","null","null"
"Quantitative analysis of platelet-activating factor in rat brain","Quantitative analysis of platelet-activating factor in rat brain","Akira Tokumura, Takashi Yotsumoto, T. Hoshikawa, Tamotsu Tanaka, Hiroaki Tsukatani","Akira Tokumura, Takashi Yotsumoto, T. Hoshikawa, Tamotsu Tanaka, Hiroaki Tsukatani","null","null","null","null","null","1992","Life Sciences","Life Sciences","Vol.51","null","303","308","eng","true","null","scientific_journal","null","null","null","null","null","null","null","null","null"
"Identification of sn-2-ω-hydroxycarboxylate-containing phospholipids in a lipid extract from bovine brain","Identification of sn-2-ω-hydroxycarboxylate-containing phospholipids in a lipid extract from bovine brain","Akira Tokumura, Tamotsu Tanaka, Takashi Yotsumoto, Hiroaki Tsukatani","Akira Tokumura, Tamotsu Tanaka, Takashi Yotsumoto, Hiroaki Tsukatani","null","Phospholipids having both a long-chain acyl (palmitoyl or stearoyl) and a short-chain hydroxycarboxylyl (C3-C9) residue were identified by GC-MS in a fraction with PAF-like activity from a bovine brain lipid extract. The hydroxyl group in the hydroxycarboxylate residue was determined to be at the omega-position by comparison of the mass spectra of the tert-butyl-dimethylsilyl derivatives of these compounds with those of synthetic hydroxybutyrate-containing phosphatidylcholines. The co-existence of short-chain hydroxycarboxylate-, monocarboxylate- and dicarboxylate-containing phospholipids in the bovine brain lipid extract suggested that these compounds were formed by peroxidation of membrane phospholipids, especially phosphatidylcholines.","Phospholipids having both a long-chain acyl (palmitoyl or stearoyl) and a short-chain hydroxycarboxylyl (C3-C9) residue were identified by GC-MS in a fraction with PAF-like activity from a bovine brain lipid extract. The hydroxyl group in the hydroxycarboxylate residue was determined to be at the omega-position by comparison of the mass spectra of the tert-butyl-dimethylsilyl derivatives of these compounds with those of synthetic hydroxybutyrate-containing phosphatidylcholines. The co-existence of short-chain hydroxycarboxylate-, monocarboxylate- and dicarboxylate-containing phospholipids in the bovine brain lipid extract suggested that these compounds were formed by peroxidation of membrane phospholipids, especially phosphatidylcholines.","null","null","1991-05","Biochemical and Biophysical Research Communications","Biochemical and Biophysical Research Communications","Vol.177","No.1","466","473","eng","true","null","scientific_journal","null","null","10.1016/0006-291X(91)92007-7","0006-291X","null","null","null","null","null"
"植物油脂中の非メチレン中断型不飽和脂肪酸:生理作用,構造解析および代謝","植物油脂中の非メチレン中断型不飽和脂肪酸:生理作用,構造解析および代謝","田中 保","Tamotsu Tanaka","null","null","null","null","null","1999","福山大学工学部紀要","福山大学工学部紀要","Vol.23","null","111","115","jpn","null","true","research_institution","null","null","null","null","null","null","null","null","null"