Thrombin and collagen induce rapid phosphorylation of a common set of cellular proteins on tyrosine in human platelets.
The ability of thrombin and collagen to induce protein-tyrosine phosphorylation in intact human platelets was assessed by using antibodies to phosphotyrosine in conjugation with immunoblots. Upon stimulation by thrombin there was an increase in the amount of protein-tyrosine phosphorylation of three bands with molecular masses of 135, 124, and 76 kDa in a time-dependent manner. The tyrosine phosphorylation in these three proteins increased in a concurrent fashion and reached a maximum level in 10 s and then a plateau or a slight decrease. Stimulation by collagen was also followed by an increase in tyrosine phosphorylation of 135- and 124-kDa proteins. Unlike stimulation by thrombin, collagen induced no obvious tyrosine phosphorylation of 76-kDa protein. The time courses for thrombin- or collagen-induced protein-tyrosine phosphorylation were similar to that for [14C] serotonin release. These results suggest that 135- and 124-kDa proteins are a common set of proteins that become phosphorylated on tyrosine residue during platelet activation.
Nakamura
S
S
Department of Biochemistry, Fukui Medical School, Japan.
Yamamura
H
H
D000911:Antibodies, Monoclonal / Q000276:immunology; D001792:Blood Platelets / Q000378:metabolism*; D015153:Blotting, Western; D003094:Collagen / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D008970:Molecular Weight; D010750:Phosphoproteins / Q000097:blood*; D010766:Phosphorylation; D019000:Phosphotyrosine; D012701:Serotonin / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology*; D013997:Time Factors; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D000911:Antibodies, Monoclonal; D010750:Phosphoproteins; D019000:Phosphotyrosine; D012701:Serotonin; D014443:Tyrosine; D003094:Collagen; D013917:Thrombin