title: Rabbit fast skeletal muscle phospholipase C. Molecular weight determination by renaturation after polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate.
abstract: Phosphoinositide specific phospholipase C from rabbit fast skeletal muscle has been enriched ca. 1,000-fold with a specific activity of 40 mumol x min-1 x mg-1. Following SDS-PAGE, renaturation of the enzyme protein in the presence of deoxycholate allowed the determination of an apparent molecular weight of 110 kDa. Gel-filtration of the native enzyme resulted in a very similar apparent molecular weight of 115 kDa, however, associated proteins of higher molecular weight were also found. Free Ca2+ concentrations needed for half-maximal activation of PtdIns(4,5)P2, PtdIns4P and PtdIns hydrolysis are 6.3 microM, 85 microM and 1.8 mM, and the Km values for these substrates 102, 340 and 937 microM, respectively.
authors.affiliation: Abteilung für Biochemie Supramolekularer Systeme, Ruhr-Universität Bochum, Germany.
authors.forename: V
authors.initials: V
authors.lastname: Windhofer
authors.forename: M
authors.initials: M
authors.lastname: Varsànyi
authors.forename: L M
authors.initials: LM
authors.lastname: Heilmeyer
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D003840:Deoxycholic Acid / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology*; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry*; D011489:Protein Denaturation; D011817:Rabbits; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry
chemical_list: D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate; D010727:Phosphoric Diester Hydrolases; D043265:Phosphatidylinositol Diacylglycerol-Lyase