Ligand-Dependent Modulation of G Protein Conformation Alters Drug Efficacy

G protein-coupled receptor (GPCR) signaling, mediated by hetero-trimeric G proteins, can be differentially controlled by agonists. At a molecular level, this is thought to occur principally via stabilization of distinct receptor conformations by individual ligands. These distinct conformations control subsequent recruitment of transducer and effector proteins. Here, we report that ligand efficacy at the calcitonin GPCR (CTR) is also correlated with ligand-dependent alterations to G protein conformation. We observe ligand-dependent differences in the sensitivity of the G protein ternary complex to disruption by GTP, due to conformational differences in the receptor-bound G protein hetero-trimer. This results in divergent agonist-dependent receptor-residency times for the hetero-trimeric G protein and different accumulation rates for downstream second messengers. This study demonstrates that factors influencing efficacy extend beyond receptor conformation(s) and expands understanding of the molecular basis for how G proteins control/influence efficacy. This has important implications for the mechanisms that underlie ligand-mediated biased agonism. [Display omitted] [Display omitted] •Ligands stabilize distinct calcitonin receptor conformations•Distinct ligand-promoted receptor complexes stabilize specific effector conformations•Differential effector conformation contributes to ligand efficacy Ligand binding to a G protein-coupled receptor not only affects the receptor conformation, but also modulates the bound G protein to shape downstream signaling.