Phospholipase A2 activity in platelets. Immuno-purification and localization of the enzyme in rat platelets.
A comparative study on phospholipase A2 activity in platelet lysates from various species was carried out using identical assay conditions with phosphatidylethanolamine as substrate. Platelet phospholipase A2, both when expressed as activity per ml blood and as specific activity in KCl extracts, was low in human, cow, pig and goat. Moderate activities, in increasing order, were found in sheep, horse and rabbit, while rats showed by far the highest activity. In the latter four species total lysate activity was recovered in 1 M KCl extracts, suggesting that the enzyme occurs either in soluble form or as a peripheral membrane-associated protein. Immune cross-reactivity with monoclonal antibodies against rat liver mitochondrial phospholipase A2 was studied in dot-blot and monoclonal antibody-Sepharose binding experiments. Only sheep and rat platelet extracts contained cross-reactive phospholipase(s) A2. Immuno-affinity chromatography of rat platelet extracts indicated virtually complete binding of total phospholipase A2 activity and yielded pure enzyme in a single purification step. Enzyme visualization by immunogold electron microscopy showed a predominant localization in the matrix of alpha-granules.
Aarsman
A J
AJ
Centre for Biomembranes and Lipid Enzymology, Utrecht, The Netherlands.
Leunissen-Bijvelt
J
J
Van den Koedijk
C D
CD
Neys
F W
FW
Verkleij
A J
AJ
Van den Bosch
H
H
D000818:Animals; D000911:Antibodies, Monoclonal; D001792:Blood Platelets / Q000201:enzymology* / Q000648:ultrastructure; D002417:Cattle; D006041:Goats; D006736:Horses; D006801:Humans; D007150:Immunohistochemistry; D010741:Phospholipases A / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification; D054467:Phospholipases A2; D011817:Rabbits; D051381:Rats; D012756:Sheep; D013045:Species Specificity; D013552:Swine
D000911:Antibodies, Monoclonal; D010741:Phospholipases A; D054467:Phospholipases A2