Go/z-biased coupling profile of the dopamine D3 receptor

Dopamine receptors are G protein coupled receptors (GPCRs) that serve as key targets for FDA-approved drugs used to treat various neuropsychiatric disorders. Notably, [~]11% of all marketed GPCR-targeting drugs act on dopamine receptors. Five GPCRs mediate the effects of endogenous dopamine and compounds used to treat Parkinsons disease, schizophrenia, and other conditions. However, on-target side effects associated with these medications highlight the need to analyze dopamine receptor signaling to design safer, more effective therapeutics. We characterized the G protein coupling of dopamine D2-like receptors and observed the striking inability of D3R to engage with Gi proteins while effectively activating Go and Gz subtypes. Applying orthogonal cell-based assays that utilize wild-type G proteins both in parental and {Delta}Gi/o/z cells, we conclusively established that D3R does not activate Gi proteins. Further analysis of Gi2:GoA and D2R:D3R chimeras revealed that this selective inability is driven by molecular determinants located within the 5 helix of Gi and the intracellular loop 2 (ICL2) of D3R. Guided by cryo-EM structures, we modeled the interface between these regions to better understand the structural basis of this selectivity. Finally, we treated hippocampal neurons in acute brain slices with selective agonists for D2R and D3R and observed marked differences in their ability to regulate endogenous adenylyl cyclase to produce cAMP, highlighting the neurophysiological significance of our findings.