GAP-43 controls the availability of secretory chromaffin granules for regulated exocytosis by stimulating a granule-associated G0.

Besides having a role in signal transduction, heterotrimeric G proteins may also be involved in membrane trafficking events as suggested by their presence in specific intracellular compartments. In chromaffin cells, G alpha 0 is associated with secretory organelles, and its activation inhibits exocytosis. Although plasma membrane-bound G proteins are activated by cell-surface receptors, the intracellular proteins controlling organelle-associated G proteins are currently unknown. GAP-43, a neuronal protein enriched in axonal growth cones and presynaptic terminals, is one possible candidate since it can directly stimulate purified G0. We have investigated the interaction of adrenal medullary GAP-43 with chromaffin granule-associated G0 and its effect on catecholamine secretion. Cytosolic and depalmitoylated membrane-extracted GAP-43 were found to stimulate guanine nucleotide binding and exchange activity in chromaffin granule membranes. In permeabilized chromaffin cells, both forms of GAP-43 blocked calcium-dependent exocytosis, and this effect was inhibited by specific antibodies against G alpha 0. A synthetic peptide corresponding to the GAP-43 domain that interacts with G0 inhibited catecholamine secretion. This effect could be selectively reversed by the COOH-terminal peptide of G alpha 0. These results indicate that GAP-43 may be an endogenous pseudoreceptor for the secretory granule-bound form of G0 and can thereby control calcium-regulated exocytosis in chromaffin cells.