Mast cells are activated by Ag-induced clustering of IgE bound to FcεRI receptors or by basic secretagogues that stimulate pertussis toxin-sensitive heterotrimeric G proteins. The cell response includes the secretion of stored molecules, such as histamine, through exocytosis and of de novo synthesized mediators, such as arachidonate metabolites. The respective roles of G proteins α and βγ subunits as well as various types of phospholipase C (PLC) in the signaling pathways elicited by basic secretagogues remain unknown. We show that a specific Ab produced against the C-terminus of Gαi3 and an anti-recombinant Gαi2 Ab inhibited, with additive effects, both exocytosis and arachidonate release from permeabilized rat peritoneal mast cells elicited by the basic secretagogues mastoparan and spermine. A specific Ab directed against Gβγ dimers prevented both secretions. Anti-PLCβ Abs selectively prevented exocytosis. The selective phosphatidylinositol 3-kinase inhibitor LY 294002 prevented arachidonate release without modifying exocytosis. Gβγ coimmunoprecipitated with PLCβ and phosphatidylinositol 3-kinase. The anti-PLCγ1 and anti-phospholipase A2 Abs selectively blocked arachidonate release. Protein tyrosine phosphorylation was inhibited by anti-Gβγ Abs, LY294002, and anti PLCγ1 Abs. These data show that the early step of basic secretagogue transduction is common to both signaling pathways, involving βγ subunits of Gi2 and Gi3 proteins. Activated Gβγ interacts, on one hand, with PLCβ to elicit exocytosis and, on the other hand, with phosphatidylinositol 3-kinase to initiate the sequential activation of PLCγ1, tyrosine kinases, and phospholipase A2, leading to arachidonate release.
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