title: Colorimetric determination of pure Mg(2+)-dependent phosphatidate phosphatase activity.
abstract: The malachite green-molybdate reagent was used for a colorimetric assay of pure Mg2(+)-dependent phosphatidate phosphatase activity. This enzyme plays a major role in fat metabolism. Enzyme activity was linear with time and protein concentration, and with the concentration of water-soluble dioctanoyl phosphatidate. The colorimetric assay was used to examine enzyme inhibition by phenylglyoxal, propranolol, and dimethyl sulfoxide. Pure enzyme and a water-soluble phosphatidate substrate were required for the assay, which should be applicable to a well-defined large-scale screen of Mg2(+)-dependent phosphatidate phosphatise inhibitors (or activators).
authors.affiliation: Department of Food Science and Rutgers Center for Lipid Research, Rutgers University, New Brunswick, NJ 08901, U.S.A.
authors.forename: Tara
authors.initials: T
authors.lastname: Havriluk
authors.forename: Fred
authors.initials: F
authors.lastname: Lozy
authors.forename: Symeon
authors.initials: S
authors.lastname: Siniossoglou
authors.forename: George M
authors.initials: GM
authors.lastname: Carman
mesh_terms: D003124:Colorimetry / Q000379:methods*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D008982:Molybdenum; D010711:Phosphatidate Phosphatase / Q000032:analysis* / Q000037:antagonists & inhibitors; D010756:Phosphoric Acids; D012394:Rosaniline Dyes; D012441:Saccharomyces cerevisiae / Q000201:enzymology
chemical_list: D010756:Phosphoric Acids; D012394:Rosaniline Dyes; C005095:malachite green; D008982:Molybdenum; D010711:Phosphatidate Phosphatase; D008274:Magnesium; C003125:phosphomolybdic acid; D004121:Dimethyl Sulfoxide