Molecular mechanisms of native ligand selectivity in catecholamine G protein-coupled receptors

Abstract Activation of G protein-coupled receptors (GPCRs) by extracellular ligands is crucial for cellular communication and modulates numerous physiological processes. Despite sharing highly similar orthosteric binding sites, catecholamine GPCRs exhibit exquisite selectivity for their native agonists, even among nearly identical chemical messengers. However, the molecular basis and evolution of receptor selectivity remain poorly understood. To elucidate the structural mechanisms of GPCR selectivity, we focus on the prototypical human β 2 -adrenergic and D 1 dopaminergic receptors, which are important drug targets and respond to the catecholamines adrenaline/noradrenaline and dopamine, respectively. Guided by structural and sequence data, we identify a small set of residues responsible for ligand selectivity. By exchanging residues at four positions in the β-adrenergic receptors and seven in the D 1 -like dopaminergic receptors, we swap the pharmacological profiles of the two subfamilies. Unexpectedly, the switch in selectivity not only involves residues interacting with the ligand, but is also controlled by regions outside the orthosteric binding site. Cryo-electron microscopy structures and computational models of the mutant receptors identify distinct molecular mechanisms contributing to selectivity in a concerted manner. Our findings provide insights into GPCR evolution and highlight strategies for protein engineering and drug design.