Phospholipase A₂ Enzymes Regulate α_IIbβ₃-mediated, but not FcγRII Receptor-mediated, pp125FAK Phosphorylation in Platelets

Summary

The α_IIbβ₃ integrin and FcγRII receptors mediate, respectively, platelet adhesion and spreading on fibrinogen and immunoglobulin (IgG) coated surfaces. Platelet adhesion to fibrinogen resulted in a partial conversion of the faster to the slower migrating (phosphorylated) form of Ca⁺²-sensitive cytosolic phospholipase A₂ (cPLA₂) but failed to trigger arachidonic acid (AA) release. Full mobility shift of cPLA₂ and a massive release of AA release were stimulated by platelet adhesion to IgG or addition of thrombin to the fibrinogen adherent platelets. IgG and thrombin induced AA production were blocked by methyl arachidonyl fluorophosphonate (MAFP), an irreversible inhibitor of cPLA₂ and the Ca⁺²-independent phospholipase A₂ (iPLA₂). In contrast, bromoenol lactone (BEL), a specific inhibitor of iPLA₂ had no effect on the release of AA. MAFP and BEL prevented pp125FAK phosphorylation and platelet spreading on fibrinogen having no effect on pp125FAK phosphorylation or platelet spreading on immobilized IgG. We conclude that α_IIbβ₃-mediated pp125FAK phosphorylation and platelet spreading on fibrinogen are regulated by PLA₂ enzymes.

Abbreviations: cPLA₂, cytosolic calcium-dependent phospholipase A2; iPLA₂, calcium-independent phospholipase A2; BSA, bovine serum albumin; mAB, monoclonal antibody; MAFP, methyl arachidonyl fluorophosphonate; pp125FAK, focal adhesion kinase; BEL, bromoenol lactone; AA, arachidonic acid.

• References

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