Clustering of Integrin α_IIb-β₃Differently Regulates Tyrosine Phosphorylation of pp72^syk, PLCγ2 and pp125^FAKin Concanavalin A-stimulated Platelets

Summary

Tyrosine phosphorylation of the non-receptor tyrosine kinases pp72^sykand pp125^FAKand of the γ2 isoform of phospholipase C (PLCγ2) in human platelets stimulated with the lectin Concanavalin A was investigated. Concanavalin A induced the rapid tyrosine phosphorylation of pp72^sykand PLCγ2 with a similar kinetics, while tyrosine phosphorylation of pp125^FAKoccurred in a later phase of platelet activation. When compared with other platelet agonists, Concanavalin A revealed to be at least as potent as collagen in inducing tyrosine phosphorylation of PLCγ2 and pp125^FAK, while tyrosine phosphorylation of pp72^sykinduced by the lectin was much stronger than that induced by thrombin or collagen. Concanavalin A-induced tyrosine phosphorylation of pp72^syk, PLCγ2 and pp125^FAKwas not dependent on platelet aggregation as it occurred normally even in the absence of sample stirring and when fibrinogen binding to integrin α_IIb-β₃was inhibited by the peptide RGDS. Tyrosine phosphorylation of pp72^syk, PLCγ2 and pp125^FAKrequired the binding of the lectin to the platelet surface, but was not observed in platelets treated with succinyl-Concanavalin A, a derivative of the lectin that interacts with the same receptors but does not promote clustering of membrane glycoproteins. Moreover, the aggregation-independent tyrosine phosphorylation of pp125^FAKand pp72^sykinduced by Concanavalin A required the expression of integrin α_IIb-β₃on the platelet surface as it was strongly inhibited in platelets from patients affected by Glanzmann thrombasthenia. By contrast, tyrosine phosphorylation of PLCγ2 occurred normally also in thrombasthenic platelets stimulated with Concanavalin A. These results demonstrate that, even in the absence of aggregation, the clustering of integrin α_IIb-β₃induced by Concanavalin A on the platelet surface directly promotes tyrosine phosphorylation of pp72^sykand pp125^FAKand provide further evidence that the oligomerization of the fibrinogen receptor promoted by its natural ligand during platelet aggregation may be responsible for the tyrosine phosphorylation of these proteins induced by physiological agonists.

• References

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