Activation of human platelets by protein kinase C antibody: role for surface phosphorylation in homeostasis.
A monoclonal antibody that inhibits protein kinase C (PKC) activity, as well as PKC pseudosubstrate inhibitory peptides, was found to cause aggregation of human platelets followed by granular secretion. Binding of this antibody to the platelet surface was demonstrated directly by flow cytometry and immunofluorescence microscopy. Assays of ecto-protein kinase activity revealed that this antibody inhibits the phosphorylation of five proteins on the platelet surface. The platelet aggregation induced by extracellular PKC inhibitors could be blocked by the addition of the membrane-impermeable phosphatase inhibitor, microcystin. Thus the inhibition of surface protein phosphorylation together with continuous dephosphorylation, namely, a decrease in the phosphorylation state of surface proteins, causes the activation of platelets. The aggregation caused by decreased surface phosphorylation appears to be initiated by the exposure of active fibrinogen-binding sites on the platelet surface, as demonstrated by the formation of fibrinogen-dependent microaggregates, as the first step in this process. We conclude that the phosphorylation of surface proteins by a platelet ecto-protein kinase C protects platelets from spontaneous aggregation and thus can play an important role in homeostatic mechanisms that maintain circulating platelets in a resting state.
Babinska
A
A
State University of New York Health Science Center at Brooklyn 11203, USA.
Ehrlich
Y H
YH
Kornecki
E
E
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D006706:Homeostasis; D006801:Humans; D008297:Male; D051379:Mice; D010766:Phosphorylation; D015539:Platelet Activation; D010974:Platelet Aggregation; D011493:Protein Kinase C / Q000276:immunology*; D011494:Protein Kinases / Q000097:blood
D000906:Antibodies; D001798:Blood Proteins; D000255:Adenosine Triphosphate; D011494:Protein Kinases; C034454:ectoprotein kinase; D011493:Protein Kinase C