Analysis of Human Dopamine D Receptor Quaternary Structure

Abstract

Background: The dopamine D receptor can form dimers/oligomers, but the molecular basis for this is poorly defined. Results: Molecular modeling, mutagenesis, and analysis of inactive state receptor crystal structures allowed assessment of models of receptor organization. Conclusion: The dopamine D receptor can assume different dimeric and a rhombic tetrameric arrangements. Significance: These findings provide understanding of the molecular basis of D receptor quaternary structure. The dopamine D receptor is a class A, rhodopsin-like G protein-coupled receptor that can form dimers and/or higher order oligomers. However, the molecular basis for production of these complexes is not well defined. Using combinations of molecular modeling, site-directed mutagenesis, and homogenous time-resolved FRET, the interfaces that allow dopamine D receptor monomers to interact were defined and used to describe likely quaternary arrangements of the receptor. These were then compared with published crystal structures of dimeric β-adrenoreceptor, μ-opioid, and CXCR4 receptors. The data indicate important contributions of residues from within each of transmembrane domains I, II, IV, V, VI, and VII as well as the intracellular helix VIII in the formation of D-D receptor interfaces within homo-oligomers and are consistent with the D receptor adopting a β-adrenoreceptor-like quaternary arrangement. Specifically, results suggest that D protomers can interact with each other via at least two distinct interfaces: the first one comprising residues from transmembrane domains I and II along with those from helix VIII and a second one involving transmembrane domains IV and V. Moreover, rather than existing only as distinct dimeric species, the results are consistent with the D receptor also assuming a quaternary structure in which two transmembrane domain I-II-helix VIII dimers interact to form a "rhombic" tetramer via an interface involving residues from transmembrane domains VI and VII. In addition, the results also provide insights into the potential contribution of molecules of cholesterol to the overall organization and potential stability of the D receptor and possibly other GPCR quaternary structures.