Selective release of phospholipase A2 and lysophosphatidylserine-specific lysophospholipase from rat platelets.
Rat platelets released phospholipase A2 and lysophospholipase upon activation with thrombin or ADP. The release of phospholipases was energy-dependent and was not in parallel with that of a known lysosomal marker enzyme, N-acetyl-beta-D-glucosaminidase. The phospholipases are derived from other granules (dense granules or alpha-granules) rather than lysosomal granules of the cells. All of the activities of both phospholipases in the cell free fraction obtained from the activated platelet reaction mixture was recovered in the supernatant after centrifugation at 105,000 X g. The degree of hydrolysis of phospholipids by the phospholipase A2 followed the order: phosphatidylethanolamine (PE) greater than phosphatidylserine (PS) greater than phosphatidylcholine (PC). Phospholipase A2 shows a broad pH optimum (greater than pH 7.0) and absolutely requires Ca2+. Lysophospholipase was specific to lysophosphatidylserine (lysoPS), and neither lysophosphatidylethanolamine (lysoPE) nor lysophosphatidylcholine (lysoPC) was hydrolyzed appreciably. Both 1-acyl- and 2-acyl-lysophosphatidylserine were equally hydrolyzed. Lysophospholipase activity shows similar pH optimum to phospholipase A2. The lysophospholipase activity was lost easily at 60 degrees C. The activity was reduced by the presence of EDTA, though low but distinct activity was observed even in the presence of EDTA. Addition of Ca2+ to the mixtures restores the full activity.
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008245:Lysophospholipase / Q000097:blood*; D008246:Lysophospholipids; D008297:Male; D017830:Octoxynol; D010718:Phosphatidylserines / Q000097:blood*; D010740:Phospholipases / Q000097:blood*; D010741:Phospholipases A / Q000097:blood*; D054467:Phospholipases A2; D011092:Polyethylene Glycols; D011489:Protein Denaturation; D051381:Rats; D011919:Rats, Inbred Strains; D012701:Serotonin / Q000097:blood; D013379:Substrate Specificity; D013917:Thrombin / Q000494:pharmacology
D008246:Lysophospholipids; D010718:Phosphatidylserines; C025059:lysophosphatidylserine; D012701:Serotonin; D011092:Polyethylene Glycols; D017830:Octoxynol; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D008245:Lysophospholipase; D013917:Thrombin