Computed atlas of the human GPCR-G protein signaling complexes

Experimental mapping of G protein-coupled receptors (GPCR)-G protein signaling coupling has illuminated hundreds of receptors, yet the coupling specificity of a large fraction of this large receptor family remains unknown, thereby preventing the development of new GPCR-targeting therapies. Here, we used AlphaFold3 (AF3) to predict the 3D structures of the human GPCRome in complex with heterotrimeric G proteins. We used experimental GPCR-G protein binding data to show that AF3 predictions significantly discriminate between positive and negative binders, and used 3D structural features to train a machine learning (ML) algorithm to predict coupling potency. Interpretation of the ML model helped discriminate universal features governing the strength of G protein coupling from those determining binding specificity. We computationally illuminated the coupling preferences of 180 non-olfactory GPCRs (non-OR) with previously unreported transduction mechanisms and experimentally validate the predicted couplings for multiple previously uncharacterized GPCRs, including QRFPR, GPR50, GPR37, GPR37L1 and GPRC5A. Our predictions established that Gi/o is the most prevalent coupling among non-OR GPCRs, which is often co-occurring with Gq/11 and, to a lesser extent, G12/13 signaling. Gs coupling is less common and restricted to specific clusters within the non-OR GPCRome phylogeny, likely due to stricter structural requirements for its binding. We also computed G protein complexes for over 400 ORs, establishing Gs as the most prevalent coupling. ORs are predicted to bind to Gs with a simpler interface compared to non-ORs, ultimately leading to energetically less stable complexes. Additionally, we predict recurrent bindings to Gq/11 and Gi/o proteins for ORs, suggesting potentially novel ORs signaling mechanisms. We exploited the GPCRome coupling atlas to interpret healthy and cancer expression data, revealing the coupling of most GPCR-G protein co-expressed pairs. This analysis highlights a richer coupling repertoire in healthy tissues compared to cancer, likely reflecting the high signaling requirements of specialized normal cell functions, which are lost in most cancer cells due to their de-differentiated state or under cancer selection processes. In summary, this study provides the first computational 3D atlas of the human GPCR-G protein transductome, thereby illuminating the signaling mechanisms of neglected GPCR classes and providing the basis for interpreting omics datasets from a myriad of pathological conditions, thus enabling the development of novel precision therapeutics.