Agonist-induced formation of unproductive receptor-G12 complexes

Abstract

G proteins are activated when they associate with G protein-coupled receptors (GPCRs), often in response to agonist-mediated receptor activation. It is generally thought that agonist-induced receptor-G protein association necessarily promotes G protein activation and, conversely, that activated GPCRs do not interact with G proteins that they do not activate. Here we show that GPCRs can form agonist-dependent complexes with G proteins that they do not activate. Using cell-based bioluminescence resonance energy transfer (BRET) and luminescence assays we find that vasopressin V₂ receptors (V₂R) associate with both G_s and G₁₂ heterotrimers when stimulated with the agonist arginine vasopressin (AVP). However, unlike V₂R-G_s complexes, V₂R-G₁₂ complexes are not destabilized by guanine nucleotides and do not promote G₁₂ activation. Activating V₂R does not lead to signaling responses downstream of G₁₂ activation, but instead inhibits basal G₁₂-mediated signaling, presumably by sequestering G₁₂ heterotrimers. Overexpressing G₁₂ inhibits G protein receptor kinase (GRK) and arrestin recruitment to V₂R and receptor internalization. Formyl peptide (FPR1 and FPR2) and Smoothened (Smo) receptors also form complexes with G₁₂ that are insensitive to nucleotides, suggesting that unproductive GPCR-G₁₂ complexes are not unique to V₂R. These results indicate that agonist-dependent receptor-G protein association does not always lead to G protein activation and may in fact inhibit G protein activation.

Keywords: G protein-coupled receptor; GPCR; arrestin; ternary complex.