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Start Over Author "Chizu Aso" Topic 2-arachidonoylglycerol
4 results on '"Chizu Aso"'

1. Enzymatic characterization of recombinant rat DDHD2: a soluble diacylglycerol lipase

Author

Hideru Obinata, Chizu Aso, Akimitsu Konishi, Noriyasu Ohshima, Kazuaki Tatei, Takashi Izumi, and Mari Araki

Subjects
0301 basic medicine, Diacylglycerol lipase, 2-Arachidonoylglycerol, Arachidonic Acids, CHO Cells, URB602, Biochemistry, Glycerides, Substrate Specificity, 03 medical and health sciences, chemistry.chemical_compound, Cricetulus, 0302 clinical medicine, Phospholipase A1, Cricetinae, Animals, Lipase, Molecular Biology, Diacylglycerol kinase, Lipoprotein lipase, biology, Biphenyl Compounds, General Medicine, Recombinant Proteins, Rats, Monoacylglycerol lipase, Lipoprotein Lipase, 030104 developmental biology, chemistry, biology.protein, 030217 neurology & neurosurgery, Endocannabinoids
Abstract

DDHD2 has been reported to exhibit phospholipase A1, triacylglycerol (TG) lipase and diacylglycerol (DG) lipase activities. However, the detailed enzymatic properties of DDHD2 have not yet been elucidated. In the current study, the substrate specificity of DDHD2 towards DG, TG and phosphatidic acid (PA) has been examined using highly purified recombinant rat DDHD2 (rDDHD2) with a liquid chromatography mass spectrometer. The k cat/Km value for DG (18:0/20:4) was much higher than those for TG (18:1/18:1/18:1), and PA (18:0/20:4) in the presence of sodium deoxycholate. The enzyme activity of rDDHD2 towards DG (18:0/20:4) was highest among all of the substrates tested. In addition, rDDHD2 was highly specific to DG substrates with a polyunsaturated fatty acid at their sn-2 position. The levels of 2-arachidonoylglycerol (2-AG) in CHO cells were quantified by gas chromatography-tandem mass spectrometry, showing that CHO cells expressing recombinant rDDHD2 contained higher levels of 2-AG when cells were treated with a monoacylglycerol lipase inhibitor, URB602. These results therefore support the idea that DDHD2 functions as a DG lipase in vivo and produces 2-AG.

Published
2016
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2. Protein purification and cloning of diacylglycerol lipase from rat brain

Author

Hideru Obinata, Chizu Aso, Akimitsu Konishi, Kazuaki Tatei, Tohko Hirano, Koji Kishimoto, Takashi Izumi, Mari Araki, Mikiko Kishi, Noriyasu Ohshima, Hiroyuki Sugimoto, and Fumio Goto

Subjects
0301 basic medicine, Diacylglycerol lipase, 2-Arachidonoylglycerol, Nerve Tissue Proteins, Arachidonic Acids, CHO Cells, Biochemistry, Hippocampus, Gene Expression Regulation, Enzymologic, Glycerides, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Cricetulus, Phospholipase A1, Protein Domains, Cricetinae, Protein purification, Animals, Lipase, Cloning, Molecular, Molecular Biology, Diacylglycerol kinase, Neurons, Lipoprotein lipase, biology, Chinese hamster ovary cell, Regular Papers, General Medicine, Rats, Lipoprotein Lipase, 030104 developmental biology, chemistry, biology.protein, Neuroglia, 030217 neurology & neurosurgery, Endocannabinoids
Abstract

Diacylglycerol (DG) lipase, which hydrolyses 1-stearoyl-2-arachidonyl-sn-glycerol to produce an endocannabinoid, 2-arachidonoylglycerol, was purified from the soluble fraction of rat brain lysates. DG lipase was purified about 1,200-fold by a sequential column chromatographic procedure. Among proteins identified by mass spectrometry analysis in the partially purified DG lipase sample, only DDHD domain containing two (DDHD2), which was formerly regarded as a phospholipase A1, exhibited significant DG lipase activity. Rat DDHD2 expressed in Chinese hamster ovary cells showed similar enzymatic properties to partially purified DG lipase from rat brain. The source of DG lipase activity in rat brain was immunoprecipitated using anti-DDHD2 antibody. Thus, we concluded that the DG lipase activity in the soluble fraction of rat brain is derived from DDHD2. DDHD2 is distributed widely in the rat brain. Immunohistochemical analysis revealed that DDHD2 is expressed in hippocampal neurons, but not in glia.

Published
2015

3. Enzymatic characterization of recombinant rat DDHD2: a soluble diacylglycerol lipase.

Author

Mari Araki, Noriyasu Ohshima, Chizu Aso, Akimitsu Konishi, Hideru Obinata, Kazuaki Tatei, and Takashi Izumi

Subjects
RECOMBINANT proteins, PHOSPHOLIPASES, MASS spectrometers, PROTEIN expression, LABORATORY rats
Abstract

DDHD2 has been reported to exhibit phospholipase A1, triacylglycerol (TG) lipase and diacylglycerol (DG) lipase activities. However, the detailed enzymatic properties of DDHD2 have not yet been elucidated. In the current study, the substrate specificity of DDHD2 towards DG, TG and phosphatidic acid (PA) has been examined using highly purified recombinant rat DDHD2 (rDDHD2) with a liquid chromatography mass spectrometer. The kcat/Km value for DG (18:0/20:4) was much higher than those for TG (18:1/18:1/18:1), and PA (18:0/20:4) in the presence of sodium deoxycholate. The enzyme activity of rDDHD2 towards DG (18:0/20:4) was highest among all of the substrates tested. In addition, rDDHD2 was highly specific to DG substrates with a polyunsaturated fatty acid at their sn−2 position. The levels of 2-arachidonoylglycerol (2-AG) in CHO cells were quantified by gas chromatography-tandem mass spectrometry, showing that CHO cells expressing recombinant rDDHD2 contained higher levels of 2-AG when cells were treated with a monoacylglycerol lipase inhibitor, URB602. These results therefore support the idea that DDHD2 functions as a DG lipase in vivo and produces 2-AG. [ABSTRACT FROM AUTHOR]

Published
2016
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4. Protein purification and cloning of diacylglycerol lipase from rat brain.

Author

Chizu Aso, Mari Araki, Noriyasu Ohshima, Kazuaki Tatei, Tohko Hirano, Hideru Obinata, Mikiko Kishi, Koji Kishimoto, Akimitsu Konishi, Fumio Goto, Hiroyuki Sugimoto, and Takashi Izumi

Subjects
*LIPASES, *DIGLYCERIDES, *PROTEIN research, *PHOSPHOLIPASE A1, *CHO cell, *MASS spectrometry, *IMMUNOHISTOCHEMISTRY, *LABORATORY rats
Abstract

Diacylglycerol (DG) lipase, which hydrolyses 1-stearoyl-2-arachidonyl-sn-glycerol to produce an endocannabinoid, 2-arachidonoylglycerol, was purified from the soluble fraction of rat brain lysates. DG lipase was purified about 1,200-fold by a sequential column chromatographic procedure. Among proteins identified by mass spectrometry analysis in the partially purified DG lipase sample, only DDHD domain containing two (DDHD2), which was formerly regarded as a phospholipase A1, exhibited significant DG lipase activity. Rat DDHD2 expressed in Chinese hamster ovary cells showed similar enzymatic properties to partially purified DG lipase from rat brain. The source of DG lipase activity in rat brain was immunoprecipitated using anti-DDHD2 antibody. Thus, we concluded that the DG lipase activity in the soluble fraction of rat brain is derived from DDHD2. DDHD2 is distributed widely in the rat brain. Immunohistochemical analysis revealed that DDHD2 is expressed in hippocampal neurons, but not in glia. [ABSTRACT FROM AUTHOR]

Published
2016
Full Text
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