title: Protein purification and cloning of diacylglycerol lipase from rat brain.
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.
authors.affiliation: Department of Biochemistry; Department of Anesthesiology, Gunma University Graduate School of Medicine, Maebashi, Gunma 371-8511; and.
authors.forename: Chizu
authors.initials: C
authors.lastname: Aso
authors.affiliation: Department of Biochemistry;
authors.forename: Mari
authors.initials: M
authors.lastname: Araki
authors.affiliation: Department of Biochemistry; ohshima-tky@umin.ac.jp.
authors.forename: Noriyasu
authors.initials: N
authors.lastname: Ohshima
authors.affiliation: Department of Biochemistry;
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Tatei
authors.affiliation: Department of Biochemistry;
authors.forename: Tohko
authors.initials: T
authors.lastname: Hirano
authors.affiliation: Department of Biochemistry;
authors.forename: Hideru
authors.initials: H
authors.lastname: Obinata
authors.affiliation: Department of Biochemistry;
authors.forename: Mikiko
authors.initials: M
authors.lastname: Kishi
authors.affiliation: Department of Biochemistry;
authors.forename: Koji
authors.initials: K
authors.lastname: Kishimoto
authors.affiliation: Department of Biochemistry;
authors.forename: Akimitsu
authors.initials: A
authors.lastname: Konishi
authors.affiliation: Department of Anesthesiology, Gunma University Graduate School of Medicine, Maebashi, Gunma 371-8511; and.
authors.forename: Fumio
authors.initials: F
authors.lastname: Goto
authors.affiliation: Department of Biochemistry, Dokkyo Medical University School of Medicine, Mibu, Tochigi 321-0293, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sugimoto
authors.affiliation: Department of Biochemistry; takashi_izumi@gunma-u.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Izumi
mesh_terms: D000818:Animals; D001095:Arachidonic Acids / Q000235:genetics / Q000378:metabolism; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D063388:Endocannabinoids / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D005989:Glycerides / Q000235:genetics / Q000378:metabolism; D006624:Hippocampus / Q000201:enzymology*; D008071:Lipoprotein Lipase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D009457:Neuroglia / Q000201:enzymology; D009474:Neurons / Q000201:enzymology*; D000072417:Protein Domains; D051381:Rats
chemical_list: D001095:Arachidonic Acids; D063388:Endocannabinoids; D005989:Glycerides; D009419:Nerve Tissue Proteins; C094503:glyceryl 2-arachidonate; D008071:Lipoprotein Lipase
keywords: 2-arachidonoylglycerol; diacylglycerol; endocannabinoid; lipase; phospholipase