Phosphorylation of the outer surface of platelets enhances the effects of collagen on aggregation, ATP release, calcium translocation and phosphoinositide hydrolysis.

Phosphorylation of the outer surface of human platelets increases their functional responsiveness to subthreshold amounts of collagen. Collagen-stimulated platelet aggregation, release of ATP and changes in cytoplasmic ionized calcium levels were all enhanced by pretreatment with ATP plus a kinase purified from human plasma. [3H]-myo-inositol-labeled human platelets were used to investigate the possible role of phosphoinositide metabolism in mediating this enhanced responsiveness. Formation of inositol mono-, bis-, and trisphosphate in response to collagen was more pronounced in phosphorylated platelets than in control platelets. Collagen-stimulated inositol phosphate production in both control and phosphorylated platelets was completely inhibited by the addition of 20 microM indomethacin. Of a range of agents that stimulated phosphoinositide hydrolysis, only the response to collagen was enhanced by phosphorylation. The plasma protein kinase was shown to catalyse phosphorylation of the collagen receptor. These data indicate that the enhanced phosphoinositide hydrolysis resulting from phosphorylation of the collagen receptor might be linked to the increased functional responses to collagen.