Abstract
G protein–coupled receptors (GPCRs) are major players in cell communication. Although they form functional monomers, increasing evidence indicates that GPCR dimerization has a critical role in cooperative phenomena that are important for cell signal integration. However, the structural bases of these phenomena remain elusive. Here, using well-characterized receptor dimers, the metabotropic glutamate receptors (mGluRs), we show that structural changes at the dimer interface are linked to receptor activation. We demonstrate that the main dimer interface is formed by transmembrane α helix 4 (TM4) and TM5 in the inactive state and by TM6 in the active state. This major change in the dimer interface is required for receptor activity because locking the TM4-TM5 interface prevents activation by agonist, whereas locking the TM6 interface leads to a constitutively active receptor. These data provide important information on the activation mechanism of mGluRs and improve our understanding of the structural basis of the negative cooperativity observed in these GPCR dimers.
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Acknowledgements
We thank the Cisbio Company for their support in providing reagents. The intracellular calcium release and FRET experiments were performed at the ARPEGE (Pharmacology Screening-Interactome) facility, Institut de Génomique Fonctionnelle (Montpellier, France). J.L. was supported by the National Natural Science Foundation of China (grant numbers 31130028 and 31225011), the Ministry of Science and Technology (grant number 2012CB518000), the Program for Introducing Talents of Discipline to the Universities of the Ministry of Education (grant number B08029) and the Mérieux Research Grants Program of the Institut Mérieux. P.R. and J.-P.P. are supported by the Centre National de la Recherche Scientifique, the Institut National de la Santé et de la Recherche Médicale and by grants from the Agence Nationale de la Recherche (ANR-09-PIRI-0011), the Fondation pour la Recherche Médicale (Equipe FRM DEQ20130326522) and the Fondation Bettencourt Schueller. L.X. was supported by a doctoral fellowship from the French Embassy in China and X.R. by a FEBS long-term fellowship and by a Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR) BP post-doctoral fellowship.
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L.X., X.R., J.L., J.-P.P. and P.R. designed experiments; L.X. performed molecular biology, cross-linking, functional assays and FRET experiments; X.R. performed molecular modeling; P.S. performed FRET experiments; H.Z. performed molecular biology; J.L., J.-P.P. and P.R. wrote the manuscript.
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Supplementary Text and Figures
Supplementary Results and Supplementary Figures 1–13. (PDF 12218 kb)
Supplementary Video 1
Movie to illustrate the changes of the mGluR2 ECD dimer between the resting and active states (side view). (MP4 510 kb)
Supplementary Video 2
Movie to illustrate the changes of the mGluR2 ECD dimer between the resting and active states (bottom view). (MP4 954 kb)
Supplementary Video 3
Movie to illustrate the major rearrangement of the mGluR2 7TM dimer between the resting and active states (top view). (MP4 1280 kb)
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Xue, L., Rovira, X., Scholler, P. et al. Major ligand-induced rearrangement of the heptahelical domain interface in a GPCR dimer. Nat Chem Biol 11, 134–140 (2015). https://doi.org/10.1038/nchembio.1711
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DOI: https://doi.org/10.1038/nchembio.1711
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