Mammalian Ric-8A (synembryn) is a heterotrimeric Gα protein guanine nucleotide exchange factor

Abstract The activation of heterotrimeric G proteins is accomplished primarily by the guanine nucleotide exchange activity of ligand-bound G protein-coupled receptors. The existence of nonreceptor guanine nucleotide exchange factors for G proteins has also been postulated. Yeast two-hybrid screens with Gαo and Gαs as baits were performed to identify binding partners of these proteins. Two mammalian homologs of the Caenorhabditis elegans protein Ric-8 were identified in these screens: Ric-8A (Ric-8/synembryn) and Ric-8B. Purification and biochemical characterization of recombinant Ric-8A revealed that it is a potent guanine nucleotide exchange factor for a subset of Gα proteins including Gαq, Gαi1, and Gαo, but not Gαs. The mechanism of Ric-8A-mediated guanine nucleotide exchange was elucidated. Ric-8A interacts with GDP-bound Gα proteins, stimulates release of GDP, and forms a stable nucleotide-free transition state complex with the Gα protein; this complex dissociates upon binding of GTP to Gα.