Platelet-activating factor (PAF)-dependent biochemical, morphologic, and physiologic responses of human platelets: demonstration of translocation of protein kinase C associated with protein phosphorylation.
Platelet-activating factor (PAF) is a potent stimulus for platelet aggregation and secretion. PAF has been shown to stimulate the phosphatidylinositol (PI) pathway in platelets, which implies that PAF should activate protein kinase C. In this study, measurements of PI metabolites, the elevation of intracellular free calcium concentration, (Ca2+)i, the activation of protein kinase C, and the phosphorylation of platelet proteins (using a two-dimensional gel electrophoretic technique) were performed before and after the addition of 10(-8) M PAF to human platelets. These findings were correlated with morphologic changes in the platelets as determined by immunoelectron microscopic studies on the cytoskeleton and by X-ray analysis of dense bodies. The results show that PAF stimulates the production of PI metabolites and causes an increase in the membrane-associated activity of protein kinase C. These changes are accompanied by a rise in the (Ca2+)i and protein phosphorylation. The increase in protein kinase C activity reaches a maximum at approximately 60 s, a time frame that is consistent with the protein phosphorylation and the subsequent morphologic and secretory events. X-ray analysis revealed two types of dense bodies containing various amounts of calcium which appeared to be released sequentially after PAF activation. These results suggest that the protein phosphorylation that controls the physiologic events resulting from PAF activation of human platelets is catalyzed by protein kinase C.
Block
L H
LH
Department of Medicine, Kantonsspital, University of Basel, Switzerland.
Abraham
W M
WM
Groscurth
P
P
Qiao
B Y
BY
Perruchoud
A P
AP
D000199:Actins / Q000097:blood; D001692:Biological Transport; D001792:Blood Platelets / Q000378:metabolism* / Q000648:ultrastructure; D002118:Calcium / Q000097:blood; D002462:Cell Membrane / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000378:metabolism; D004075:Diglycerides / Q000097:blood; D004577:Electron Probe Microanalysis; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007295:Inositol Phosphates / Q000097:blood; D008970:Molecular Weight; D010716:Phosphatidylinositols / Q000097:blood*; D010766:Phosphorylation; D010972:Platelet Activating Factor / Q000502:physiology*; D010974:Platelet Aggregation / Q000502:physiology; D011493:Protein Kinase C / Q000097:blood*
D000199:Actins; D004075:Diglycerides; D007295:Inositol Phosphates; D010716:Phosphatidylinositols; D010972:Platelet Activating Factor; D011493:Protein Kinase C; D002118:Calcium