Abstract
The dopaminergic system, including five dopamine receptors (D1R to D5R), plays essential roles in the central nervous system (CNS); and ligands that activate dopamine receptors have been used to treat many neuropsychiatric disorders, including Parkinson’s Disease (PD) and schizophrenia. Here, we report cryo-EM structures of all five subtypes of human dopamine receptors in complex with G protein and bound to the pan-agonist, rotigotine, which is used to treat PD and restless legs syndrome. The structures reveal the basis of rotigotine recognition in different dopamine receptors. Structural analysis together with functional assays illuminate determinants of ligand polypharmacology and selectivity. The structures also uncover the mechanisms of dopamine receptor activation, unique structural features among the five receptor subtypes, and the basis of G protein coupling specificity. Our work provides a comprehensive set of structural templates for the rational design of specific ligands to treat CNS diseases targeting the dopaminergic system.
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Data availability
Density maps and structure coordinates have been deposited in the Electron Microscopy Data Bank (EMDB) and the Protein Data Bank (PDB) with accession codes EMD-35683 and 8IRR for the D1R–Gs–rotigotine complex; EMD-35684 and 8IRS for the D2R–Gi–rotigotine complex; EMD-35685 and 8IRT for the D3R–Gi–rotigotine complex; EMD-35686 and 8IRU for the D4R–Gi–rotigotine complex; EMD-35687 and 8IRV for the D5R–Gs–rotigotine complex.
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Acknowledgements
The cryo-EM data were collected at the Center of Cryo-Electron Microscopy, Shanghai Institute of Materia Medica, the Center of Cryo-Electron Microscopy, Zhejiang University, and the Cryo-Electron Microscopy Facility, Zhejiang University Medical Center/Liangzhu laboratory. This work was partially supported by the National Key R&D Programs of China (2018YFA0507002), Shanghai Municipal Science and Technology Major Project (2019SHZDZX02 and XDB08020303) to H.E.X.; the National Key Basic Research Program of China (2019YFA0508800), the Key R&D Projects of Zhejiang Province (2021C03039) and Fundamental Research Funds for the Central Universities (2019XZZX001-01-06) to Yan Z.; the Zhejiang Province Natural Science Fund for Excellent Young Scholars (LR22C050002) and the National Natural Science Foundation of China (32100959) to C.M.; the National Natural Science Foundation of China (31770796) and the National Science and Technology Major Project (2018ZX09711002) to Y.J.; grants from the NIMH Psychoactive Drug Screening Program to X.-P.H., Y.L., B.L.R., and RO1MH112205 to B.E.K. and B.L.R. The Special Research Assistant Project of Chinese Academy of Sciences to Youwen Z.
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P.X. and S.H. designed the expression constructs, purified the complexes, and prepared protein samples for the D2R–Gi, D3R–Gi, D4R–Gi, and D5R–Gs complexes for cryo-EM data collection. P.X. and S.H. performed cryo-EM grid preparation, data acquisition, structure determination, and prepared the draft of the manuscript and figures. Youwen Z. designed the constructs, prepared the protein samples, conducted cryo-EM data collection and structure determination of D1R–Gs, and participated in the preparation of supplementary figures and manuscript editing. C.M. screened the cryo-EM conditions, prepared the cryo-EM grids, collected cryo-EM images, processed the EM data of the D2R–Gi complex, and participated in the preparation of supplementary figures. P.X. built the models and refined the structures. Yumu Z. and H.L. participated in the sample preparation and screening of the D4R–Gi and the D5R–Gs complexes. Y.W. participated in the sample preparation and screening of the D1R–Gs complex. B.E.K., X.-P.H., and Y.-F.L. performed cAMP, GPCRome, Tango, and radioligand binding assays. B.E.K. compiled assay data and participated in the preparation of the manuscript. X.H. performed the docking studies. W.Y. designed the Gα constructs used for the generation of the D4R–Gi complex. Y.J. participated in the funding acquisition. Yan Z. supervised C.M. and participated in manuscript editing. B.L.R. supervised pharmacological and mutagenesis experiments and participated in manuscript writing. H.E.X. conceived and supervised the project and wrote the manuscript with P.X.
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Xu, P., Huang, S., Krumm, B.E. et al. Structural genomics of the human dopamine receptor system. Cell Res 33, 604–616 (2023). https://doi.org/10.1038/s41422-023-00808-0
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DOI: https://doi.org/10.1038/s41422-023-00808-0