Partial purification and properties of microsomal phosphatidate phosphohydrolase from rat liver.
Microsomal phosphatidate phosphohydrolase (phosphatidate phosphatase EC 3.1.3.4) was solubilized and fractionated to yield at least two distinct enzymatically active fractions. One, denoted FA, was non-specific, had a relatively high Km for phosphatidic acid and was insensitive to inhibition by diacylglycerol. The second fraction, FB, was specific for phosphatidates, had a low Km, and was inhibited, non-competitively, by diacylglycerol. FA exhibited a sigmoid substrate-activity curve. The isolated FB aggregated to particles of about 10(6) in the absence of salts and could be dissociated by the addition of monovalent cations at ionic strength 0.4-0.6 to about 2-10(5) daltons and thereby doubled its activity. Dissociation was time- and temperature-dependent. F- was inhibitory. Divalent ions were not required for the activity of FA or FB and inhibited at concentrations exceeding 1 mM.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D003840:Deoxycholic Acid; D004075:Diglycerides / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008862:Microsomes, Liver / Q000201:enzymology*; D009994:Osmolar Concentration; D010743:Phospholipids / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D012965:Sodium Chloride / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013696:Temperature
D004075:Diglycerides; D010743:Phospholipids; D003840:Deoxycholic Acid; D008345:Manganese; D012965:Sodium Chloride; D010744:Phosphoric Monoester Hydrolases; D008274:Magnesium; D002118:Calcium