Evaluation of the fatty acid selectivity of a phosphatidylinositol-specific cytosolic phospholipase C from pig and human platelets.
The fatty acid selectivity of cytosolic phospholipase C (phosphatidylinositol phosphodiesterase) from pig and human platelets towards phosphatidylinositol was evaluated. For this purpose, the relative conversion of rat liver phosphatidylinositol (enriched in stearate and arachidonate) and sheep liver phosphatidylinositol (enriched in stearate plus oleate and containing linoleate and arachidonate) was compared and, in addition, the fatty acid compositions of the diacylglycerol products were determined by gas-liquid chromatography. The cytosolic enzyme exhibited essentially complete specificity for phosphatidylinositol when choline-, ethanolamine-, serine-, or inositol-containing phospholipids labelled with [14C]stearate were tested as substrates. Similar percentage conversions of rat and sheep liver phosphatidylinositols to 1,2-diacylglycerol were found with phospholipase C from either pig or human platelets. Furthermore, the newly formed diacylglycerols and the unreacted phospholipid had fatty acid compositions which were very similar to the corresponding substrates. These results suggest that the phospholipase C from isolated platelet cytosol is highly selective towards phosphatidylinositol, but not with respect to the fatty acid composition of naturally occurring phosphatidylinositol. They also suggest that any preferential release of arachidonoyl diacylglycerol in stimulated human platelets is more likely controlled by compartmentation of the corresponding phosphatidylinositol precursor within platelet membranes and its availability, rather than directly by a marked enzyme preference for arachidonate-containing species.
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology; D004075:Diglycerides / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D006801:Humans; D010716:Phosphatidylinositols / Q000378:metabolism*; D010740:Phospholipases / Q000097:blood*; D010743:Phospholipids / Q000378:metabolism; D012756:Sheep; D013552:Swine; D010738:Type C Phospholipases / Q000097:blood*
D004075:Diglycerides; D005227:Fatty Acids; D010716:Phosphatidylinositols; D010743:Phospholipids; D010740:Phospholipases; D010738:Type C Phospholipases