Lipid phosphate phosphatases from Saccharomyces cerevisiae.
DPP1-encoded and LPP1-encoded lipid phosphate phosphatases are integral membrane proteins in the yeast Saccharomyces cerevisiae. They catalyze the Mg(2+)-independent dephosphorylation of bioactive lipid phosphate molecules such as diacylglycerol pyrophosphate and phosphatidate. These enzymes possess a three-domain lipid phosphatase motif that is localized to the hydrophilic surface of the membrane. The lipid phosphate phosphatase activities of DPP1-encoded and LPP1-encoded enzymes are measured by following the release of water-soluble radioactive inorganic phosphate from chloroform-soluble radioactive lipid phosphate substrate following a chloroform/methanol/water phase partition. The DPP1-encoded enzyme, commonly referred to as diacylglycerol pyrophosphate phosphatase, is purified from wild-type S. cerevisiae membranes by detergent solubilization with Triton X-100 followed by chromatography with DEAE-cellulose (DE53), Affi-Gel blue, hydroxylapatite, and Mono Q. The purification scheme yields an essentially homogeneous enzyme preparation that is stable for several years upon storage at -80 degrees . The properties of the DPP1-encoded and LPP1-encoded lipid phosphate phosphatase enzymes are summarized.
Carman
George M
GM
Department of Food Science, Rutgers University, New Brunswick, New Jersey, USA.
Wu
Wen-I
WI
D002462:Cell Membrane / Q000201:enzymology; D002848:Chromatography, DEAE-Cellulose / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D004795:Enzyme Stability; D005615:Freezing; D007700:Kinetics; D008055:Lipids / Q000502:physiology; D008861:Microsomes / Q000201:enzymology; D010711:Phosphatidate Phosphatase / Q000302:isolation & purification / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity; D014617:Vacuoles / Q000201:enzymology
D008055:Lipids; D010761:Phosphorus Radioisotopes; D029701:Saccharomyces cerevisiae Proteins; C112948:lipid phosphate phosphatase; D010711:Phosphatidate Phosphatase