Abstract
SB269652 is to our knowledge the first drug-like allosteric modulator of the dopamine D2 receptor (D2R), but it contains structural features associated with orthosteric D2R antagonists. Using a functional complementation system to control the identity of individual protomers within a dimeric D2R complex, we converted the pharmacology of the interaction between SB269652 and dopamine from allosteric to competitive by impairing ligand binding to one of the protomers, indicating that the allostery requires D2R dimers. Additional experiments identified a 'bitopic' pose for SB269652 extending from the orthosteric site into a secondary pocket at the extracellular end of the transmembrane (TM) domain, involving TM2 and TM7. Engagement of this secondary pocket was a requirement for the allosteric pharmacology of SB269652. This suggests a new mechanism whereby a bitopic ligand binds in an extended pose on one G protein–coupled receptor protomer to allosterically modulate the binding of a ligand to the orthosteric site of a second protomer.
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Acknowledgements
We thank TGR BioSciences for generously providing the SureFire ERK1/2 kits. A.C. and P.M.S. are Principal Research Fellows of the National Health and Medical Research Council (NHMRC) of Australia. J.R.L. is a Monash University Larkins Fellow and an R.D. Wright Biomedical Career Development Fellow of the NHMRC. This work was funded in part by NHMRC Program Grant no. APP1055134 (A.C. and P.M.S.), Project Grant no. APP1011920 (J.R.L.), Project Grant APP1049564 (J.R.L. and B.C.) and Australian Research Council Discovery Grant no. DP110100687 (P.J.S. and A.C.). J.R.L. acknowledges the financial support of the Netherlands Organization for Scientific Research (NWO VENI Grant 863.09.018). This work was supported in part by US National Institutes of Health grants DA022413, MH54137 (J.A.J.) and DA023694 (L.S.) and the Lieber Center for Schizophrenia Research and Treatment (J.A.J.). A. Stewart is thanked for technical assistance.
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J.R.L. conceived and supervised the project, generated receptor mutants and cell lines, performed data analysis, performed radioligand binding and functional assays and wrote the manuscript. P.D. performed complementation assay experiments, generated receptor constructs and wrote the manuscript. J.S. synthesized aripiprazole, SB269652 and its derivatives and wrote the corresponding experimental section. C.J.D.-J. performed radioligand binding and functional assays, generated mutant receptors and cell lines and performed data analysis. S.D. performed functional assays. M.M. conducted docking and homology modeling. L.S. conducted and supervised docking and homology modeling and wrote the manuscript. L.L. planned mutagenesis experiments. P.J.S. planned and supervised chemical synthesis and wrote the manuscript. B.C. planned and supervised chemical synthesis and wrote the manuscript. P.M.S. supervised the project and wrote the manuscript. J.A.J. supervised the project and wrote the manuscript. A.C. conceived and supervised the project, performed data analysis and wrote the manuscript.
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A.C. and P.M.S. have a research contract with Servier, France. A.C. has had recent consultancies with Johnson and Johnson (USA) and XOMA (USA) and is a Scientific Advisory Board member for Audeo, Australia.
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Lane, J., Donthamsetti, P., Shonberg, J. et al. A new mechanism of allostery in a G protein–coupled receptor dimer. Nat Chem Biol 10, 745–752 (2014). https://doi.org/10.1038/nchembio.1593
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DOI: https://doi.org/10.1038/nchembio.1593
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