Protein tyrosine phosphorylation in human platelets induced by interaction between glycoprotein Ib and von Willebrand factor.
The interaction between von Willebrand factor (vWF) and glycoprotein Ib (GPIb) induced by ristocetin or botrocetin resulted in associated platelet aggregation, protein tyrosine phosphorylation (PTP) of a 64 kDa protein, as detected by a monoclonal antibody against phosphotyrosine (PY-20), and intracellular Ca2+ elevation that is largely dependent upon Ca2+ influx in human platelets. It is of interest that 75-80, 97 and 125 kDa proteins which are strongly tyrosine-phosphorylated in platelet activation induced by thrombin and other agonists were not detected. Neither vWF nor a coaggregating agent (ristocetin or botrocetin) alone induced aggregation, [Ca2+]i elevation or the 64 kDa PTP. NMC-4, an antibody which inhibits both ristocetin- or botrocetin-induced vWF binding to GPIb, abolished the appearance of the 64 kDa PTP as well as other responses, suggesting that it is specifically induced by the GPIb-vWF interaction. Aspirin, or ONO-3708, a competitive inhibitor of thromboxane A2, did not modify the 64 kDa PTP, while [Ca2+]i elevation was moderately suppressed. Depletion of extracellular Ca2+ or RGD peptides suppressed neither the 64 kDa PTP nor aggregation. H-7, a protein kinase C inhibitor, did not inhibit the 64 kDa PTP, while staurosporine, a potent protein kinase inhibitor, inhibited the 64 kDa PTP and Ca2+ influx, but not aggregation, in a dose-dependent manner. It is suggested that the 64 kDa PTP is associated with platelet aggregation induced by the interaction between GPIb and vWF.
Ozaki
Y
Y
Department of Clinical and Laboratory Medicine, Yamanashi Medical University, Japan.
Satoh
K
K
Yatomi
Y
Y
Miura
S
S
Fujimura
Y
Y
Kume
S
S
D000470:Alkaloids / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000097:blood; D003435:Crotalid Venoms / Q000494:pharmacology; D011464:Epoprostenol / Q000494:pharmacology; D006801:Humans; D010750:Phosphoproteins / Q000097:blood*; D010766:Phosphorylation; D019000:Phosphotyrosine; D010974:Platelet Aggregation / Q000187:drug effects; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D012310:Ristocetin / Q000494:pharmacology; D019311:Staurosporine; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000097:blood; D014841:von Willebrand Factor / Q000378:metabolism* / Q000494:pharmacology
D000470:Alkaloids; D003435:Crotalid Venoms; D010750:Phosphoproteins; D010980:Platelet Membrane Glycoproteins; D014841:von Willebrand Factor; D012310:Ristocetin; D019000:Phosphotyrosine; D014443:Tyrosine; C030961:botrocetin; D011464:Epoprostenol; D019311:Staurosporine; D002118:Calcium