Phosphoinositide 3-kinase gamma and p85/phosphoinositide 3-kinase in platelets - Relative activation by thrombin receptor or beta-phorbol myristate acetate and roles in promoting the ligand-binding function of alpha(IIb)beta(3) integrin

Platelets exposed to thrombin or thrombin receptor agonist peptide (SFLLRN) activate phospholipase C and protein kinase C (PKC), and accumulate 3-phosphorylated phosphoinositides (S-PPI) as a function of the activation and relocalization of two cytoskeletally-associated phosphoinositide 3-kinases (PI 3-K): p85/PI 3-K and PI 3-K gamma. We now report that exposure of platelets to PKC-activating beta-phorbol myristate acetate (beta PMA) does not stimulate PI 3-K gamma, but rather stimulates p85/PI 3-K, which associates with the cytoskeleton. Wortmannin is an inhibitor of both PI 3-Ks, known to act with more potency on p85/PI 3-K, beta PMA-stimulated 3-PPI accumulation is more sensitive to wortmannin (IC50 = 1.3 nM) than is SFLLRN- or thrombin-stimulatad 3-PPI accumulation (IC50 = 10 nM). The activity of p85/PI 3-K in immunoprecipitates or in cytoskeletal fractions is inhibited more potently by exposure of platelets to wortmannin than is the activity of PI 3-K gamma. beta PMA or SFLLRN promotes the conversion of platelet integrin alpha(IIb)beta(3) into a fibrinogen-binding form required for platelet aggregation. Activation of alpha(IIb)beta(3) in response to beta PMA or SFLLRN is inhibited by wortmannin with an IC50 of 1 nM in each case. Wortmannin inhibits neither activation of alpha(IIb)beta(3) by ligand-induced binding site antibody (anti-LIBS6 Fab) nor anti-LIBS6 Fab-induced platelet aggregation in the presence of fibrinogen, indicating that this type of ''outside-in'' signaling by alpha(IIb)beta(3) is largely PI 3-K-independent. We conclude that p85/PI 3-K, in preference to PI 3-K gamma, contributes to activation of alpha(IIb)beta(3) when the thrombin receptor or PKC is stimulated.