G-protein-coupled receptors (GPCRs) are key cell-surface proteins that transduce external environmental cues into biochemical signals across the membrane. GPCRs are intrinsically allosteric proteins; they interact via spatially distinct yet conformationally linked domains with both endogenous and exogenous proteins, nutrients, metabolites, hormones, small molecules and biological agents. Here we explore recent high-resolution structural studies, which are beginning to unravel the atomic details of allosteric transitions that govern GPCR biology, as well as highlighting how the wide diversity of druggable allosteric sites across these receptors present opportunities for developing new classes of therapeutics.
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Changeux, J. P. & Christopoulos, A. Allosteric modulation as a unifying mechanism for receptor function and regulation. Cell 166, 1084–1102 (2016). This review highlights the key principles of allostery as common elements across multiple receptor superfamilies
Hauser, A. S., Attwood, M. M., Rask-Andersen, M., Schioth, H. B. & Gloriam, D. E. Trends in GPCR drug discovery: new agents, targets and indications. Nat. Rev. Drug Discov. 16, 829–842 (2017).
Tesmer, J. J. Hitchhiking on the heptahelical highway: structure and function of 7TM receptor complexes. Nat. Rev. Mol. Cell Biol. 17, 439–450 (2016).
Tehan, B. G., Bortolato, A., Blaney, F. E., Weir, M. P. & Mason, J. S. Unifying family A GPCR theories of activation. Pharmacol. Ther. 143, 51–60 (2014).
Yuan, S., Filipek, S., Palczewski, K. & Vogel, H. Activation of G-protein-coupled receptors correlates with the formation of a continuous internal water pathway. Nat. Commun. 5, 4733 (2014).
Choe, H. W. et al. Crystal structure of metarhodopsin II. Nature 471, 651–655 (2011).
Kent, R. S., De Lean, A. & Lefkowitz, R. J. A quantitative analysis of beta-adrenergic receptor interactions: resolution of high and low affinity states of the receptor by computer modeling of ligand binding data. Mol. Pharmacol. 17, 14–23 (1980).
Eddy, M. T., Didenko, T., Stevens, R. C. & Wuthrich, K. β2-adrenergic receptor conformational response to fusion protein in the third intracellular loop. Structure 24, 2190–2197 (2016).
Van Eps, N. et al. Conformational equilibria of light-activated rhodopsin in nanodiscs. Proc. Natl Acad. Sci. USA 114, E3268–E3275 (2017).
Manglik, A. et al. Structural insights into the dynamic process of β2-adrenergic receptor signaling. Cell 161, 1101–1111 (2015).
Ye, L., Van Eps, N., Zimmer, M., Ernst, O. P. & Prosser, R. S. Activation of the A2A adenosine G-protein-coupled receptor by conformational selection. Nature 533, 265–268 (2016). These NMR studies on rhodopsin 9 , β 2 AR 10 and A 2A R 11 demonstrate the highly dynamic nature of the GPCR conformational landscape
Staus, D. P. et al. Allosteric nanobodies reveal the dynamic range and diverse mechanisms of G-protein-coupled receptor activation. Nature 535, 448–452 (2016). This study reports two distinct inactive state conformations and provides direct structural evidence for a dynamic range of both active and inactive GPCR states
Hino, T. et al. G-protein-coupled receptor inactivation by an allosteric inverse-agonist antibody. Nature 482, 237–240 (2012).
Carpenter, B. & Tate, C. G. Active state structures of G protein-coupled receptors highlight the similarities and differences in the G protein and arrestin coupling interfaces. Curr. Opin. Struct. Biol. 45, 124–132 (2017).
Shihoya, W. et al. Activation mechanism of endothelin ETB receptor by endothelin-1. Nature 537, 363–368 (2016).
Huang, J., Chen, S., Zhang, J. J. & Huang, X. Y. Crystal structure of oligomeric β1-adrenergic G protein-coupled receptors in ligand-free basal state. Nat. Struct. Mol. Biol. 20, 419–425 (2013).
Leslie, A. G., Warne, T. & Tate, C. G. Ligand occupancy in crystal structure of β1-adrenergic G protein-coupled receptor. Nat. Struct. Mol. Biol. 22, 941–942 (2015).
Christopoulos, A. et al. International Union of Basic and Clinical Pharmacology. XC. Multisite pharmacology: recommendations for the nomenclature of receptor allosterism and allosteric ligands. Pharmacol. Rev. 66, 918–947 (2014). This document provides the official nomenclature for ligand classification
Hori, T. et al. Na+-mimicking ligands stabilize the inactive state of leukotriene B4 receptor BLT1. Nat. Chem. Biol. 14, 262–269 (2018).
Kruse, A. C. et al. Activation and allosteric modulation of a muscarinic acetylcholine receptor. Nature 504, 101–106 (2013). This landmark study reports the first crystal structure of an active-state agonist-bound GPCR in complex with a synthetic allosteric modulator
DeVree, B. T. et al. Allosteric coupling from G protein to the agonist-binding pocket in GPCRs. Nature 535, 182–186 (2016). This paper provides direct structural evidence of the reciprocal allosteric coupling between G-protein and agonist-binding sites on a GPCR
Dror, R. O. et al. Structural basis for modulation of a G-protein-coupled receptor by allosteric drugs. Nature 503, 295–299 (2013). This key computational biology study highlights the utility of molecular dynamics for identifying mechanisms underlying the actions of allosteric modulation at a GPCR
Cheng, R. K. Y. et al. Structural insight into allosteric modulation of protease-activated receptor 2. Nature 545, 112–115 (2017).
Tan, Q. et al. Structure of the CCR5 chemokine receptor–HIV entry inhibitor maraviroc complex. Science 341, 1387 (2013).
Huma, Z. E. et al. Key determinants of selective binding and activation by the monocyte chemoattractant proteins at the chemokine receptor CCR2. Sci. Signal. 10, eaai8529 (2017).
Zheng, Y. et al. Structure of CC chemokine receptor 5 with a potent chemokine antagonist reveals mechanisms of chemokine recognition and molecular mimicry by HIV. Immunity 46, 1005–1017.e1005 (2017).
Wu, H. et al. Structure of a class C GPCR metabotropic glutamate receptor 1 bound to an allosteric modulator. Science 344, 58 (2014).
Dore, A. S. et al. Structure of class C GPCR metabotropic glutamate receptor 5 transmembrane domain. Nature 511, 557–562 (2014).
Christopher, J. A. et al. Fragment and structure-based drug discovery for a class C GPCR: Discovery of the mGlu5 negative allosteric modulator HTL14242 (3-chloro-5-[6-(5-fluoropyridin-2-yl)pyrimidin-4-yl]benzonitrile). J. Med. Chem. 58, 6653–6664 (2015).
Christopher, J. A. et al. Structure-based optimization strategies for G protein-coupled receptor (GPCR) allosteric modulators: a case study from analyses of new metabotropic glutamate receptor 5 (mGlu5) X-ray structures. J. Med. Chem. (2018).
Wang, C. et al. Structural basis for Smoothened receptor modulation and chemoresistance to anticancer drugs. Nat. Commun. 5, 4355 (2014).
Wootten, D. et al. Polar transmembrane interactions drive formation of ligand-specific and signal pathway-biased family B G protein-coupled receptor conformations. Proc. Natl. Acad. Sci. USA 110, 5211–5216 (2013).
Hollenstein, K. et al. Structure of class B GPCR corticotropin-releasing factor receptor 1. Nature 499, 438–443 (2013).
Jazayeri, A. et al. Extra-helical binding site of a glucagon receptor antagonist. Nature 533, 274–277 (2016).
Zhang, H. et al. Structure of the full-length glucagon class B G-protein-coupled receptor. Nature 546, 259–264 (2017).
Song, G. et al. Human GLP-1 receptor transmembrane domain structure in complex with allosteric modulators. Nature 546, 312–315 (2017).
Zhang, D. et al. Two disparate ligand-binding sites in the human P2Y1 receptor. Nature 520, 317–321 (2015).
Srivastava, A. et al. High-resolution structure of the human GPR40 receptor bound to allosteric agonist TAK-875. Nature 513, 124–127 (2014).
Lu, J. et al. Structural basis for the cooperative allosteric activation of the free fatty acid receptor GPR40. Nat. Struct. Mol. Biol. 24, 570–577 (2017).
Robertson, N. et al. Structure of the complement C5a receptor bound to the extra-helical antagonist NDT9513727. Nature 553, 111–114 (2018).
Zheng, Y. et al. Structure of CC chemokine receptor 2 with orthosteric and allosteric antagonists. Nature 540, 458–461 (2016).
Oswald, C. et al. Intracellular allosteric antagonism of the CCR9 receptor. Nature 540, 462–465 (2016).
Liu, X. et al. Mechanism of intracellular allosteric β2AR antagonist revealed by X-ray crystal structure. Nature 548, 480–484 (2017).
Nolte, W. M. et al. A potentiator of orthosteric ligand activity at GLP-1R acts via covalent modification. Nat. Chem. Biol. 10, 629–631 (2014).
van der Westhuizen, E. T., Valant, C., Sexton, P. M. & Christopoulos, A. Endogenous allosteric modulators of G protein–coupled receptors. J. Pharmacol. Exp. Ther. 353, 246–260 (2015).
Hay, D. L. & Pioszak, A. A. Receptor activity-modifying proteins (RAMPs): new insights and roles. Annu. Rev. Pharmacol. Toxicol. 56, 469–487 (2016).
Pert, C. B. & Snyder, S. H. Properties of opiate-receptor binding in rat brain. Proc. Natl Acad. Sci. USA 70, 2243–2247 (1973).
Katritch, V. et al. Allosteric sodium in class A GPCR signaling. Trends Biochem. Sci. 39, 233–244 (2014).
Liu, W. et al. Structural basis for allosteric regulation of GPCRs by sodium ions. Science 337, 232–236 (2012). This 1.80 Å resolution structure of the A 2A R was the first to directly identify the location of a conserved allosteric sodium site on a GPCR
Miller-Gallacher, J. L. et al. The 2.1 Å resolution structure of cyanopindolol-bound β1-adrenoceptor identifies an intramembrane Na+ ion that stabilises the ligand-free receptor. PLoS One 9, e92727 (2014).
Hanson, M. A. et al. A specific cholesterol binding site is established by the 2.8 Å structure of the human β2-adrenergic receptor. Structure 16, 897–905 (2008).
Gimpl, G. Interaction of G protein coupled receptors and cholesterol. Chem. Phys. Lipids 199, 61–73 (2016).
Manna, M. et al. Mechanism of allosteric regulation of β2-adrenergic receptor by cholesterol. eLife 5, e18432 (2016).
Cherezov, V. et al. High-resolution crystal structure of an engineered human β2-adrenergic G protein-coupled receptor. Science 318, 1258–1265 (2007).
Hua, T. et al. Crystal structures of agonist-bound human cannabinoid receptor CB1. Nature 547, 468–471 (2017).
Zhang, J. et al. Agonist-bound structure of the human P2Y12 receptor. Nature 509, 119–122 (2014).
Huang, P. et al. Cellular cholesterol directly activates Smoothened in Hedgehog signaling. Cell 166, 1176–1187 (2016).
Byrne, E. F. X. et al. Structural basis of Smoothened regulation by its extracellular domains. Nature 535, 517–522 (2016).
Luchetti, G. et al. Cholesterol activates the G-protein coupled receptor Smoothened to promote Hedgehog signaling. eLife 5, e20304 (2016).
Manglik, A., Kobilka, B. K. & Steyaert, J. Nanobodies to study G Protein-coupled receptor structure and function. Annu. Rev. Pharmacol. Toxicol. 57, 19–37 (2017).
Rasmussen, S. G. et al. Crystal structure of the β2 adrenergic receptor–Gs protein complex. Nature 477, 549–555 (2011).
Shukla, A. K. et al. Structure of active β-arrestin-1 bound to a G-protein-coupled receptor phosphopeptide. Nature 497, 137–141 (2013).
Zhang, Y. et al. Cryo-EM structure of the activated GLP-1 receptor in complex with a G protein. Nature 546, 248–253 (2017).
Liang, Y. L. et al. Phase-plate cryo-EM structure of a class B GPCR-G-protein complex. Nature 546, 118–123 (2017).
Gurbel, P. A. et al. Cell-penetrating pepducin therapy targeting PAR1 in subjects with coronary artery disease. Arterioscler. Thromb. Vasc. Biol. 36, 189–197 (2016).
Kahsai, A. W. et al. Conformationally selective RNA aptamers allosterically modulate the β2-adrenoceptor. Nat. Chem. Biol. 12, 709–716 (2016).
Hutchings, C. J., Koglin, M., Olson, W. C. & Marshall, F. H. Opportunities for therapeutic antibodies directed at G-protein-coupled receptors. Nat. Rev. Drug Discov. 16, 661 (2017).
Maguire, M. E., Van Arsdale, P. M. & Gilman, A. G. An agonist-specific effect of guanine nucleotides on binding to the beta adrenergic receptor. Mol. Pharmacol. 12, 335–339 (1976).
De Lean, A., Stadel, J. M. & Lefkowitz, R. J. A ternary complex model explains the agonist-specific binding properties of the adenylate cyclase-coupled beta-adrenergic receptor. J. Biol. Chem. 255, 7108–7117 (1980).
Gurevich, V. V., Pals-Rylaarsdam, R., Benovic, J. L., Hosey, M. M. & Onorato, J. J. Agonist-receptor-arrestin, an alternative ternary complex with high agonist affinity. J. Biol. Chem. 272, 28849–28852 (1997).
Smith, J. S., Lefkowitz, R. J. & Rajagopal, S. Biased signalling: from simple switches to allosteric microprocessors. Nat. Rev. Drug Discov. 17, 243–260 (2018).
Edelstein, S. J. & Changeux, J. P. Biased allostery. Biophys. J. 111, 902–908 (2016).
Flock, T. et al. Selectivity determinants of GPCR–G-protein binding. Nature 545, 317–322 (2017).
Gurevich, E. V., Tesmer, J. J., Mushegian, A. & Gurevich, V. V. G protein-coupled receptor kinases: more than just kinases and not only for GPCRs. Pharmacol. Ther. 133, 40–69 (2012).
He, Y. et al. Molecular assembly of rhodopsin with G protein-coupled receptor kinases. Cell Res. 27, 728–747 (2017).
Komolov, K. E. et al. Structural and functional analysis of a β2-adrenergic receptor complex with GRK5. Cell 169, 407–421 (2017).
Gurevich, V. V. & Gurevich, E. V. The molecular acrobatics of arrestin activation. Trends Pharmacol. Sci. 25, 105–111 (2004).
Shukla, A. K. et al. Visualization of arrestin recruitment by a G-protein-coupled receptor. Nature 512, 218–222 (2014).
Kim, Y. J. et al. Crystal structure of pre-activated arrestin p44. Nature 497, 142–146 (2013).
Kang, Y. et al. Crystal structure of rhodopsin bound to arrestin by femtosecond X-ray laser. Nature 523, 561–567 (2015).
Zhou, X. E. et al. Identification of phosphorylation codes for arrestin recruitment by G protein-coupled receptors. Cell 170, 457–469 (2017).
Kim, J. et al. Functional antagonism of different G protein-coupled receptor kinases for β-arrestin-mediated angiotensin II receptor signaling. Proc. Natl Acad. Sci. USA 102, 1442–1447 (2005).
Thomsen, A. R. et al. GPCR–G protein–β-arrestin super-complex mediates sustained G protein signaling. Cell 166, 907–919 (2016).
Wacker, D. et al. Structural features for functional selectivity at serotonin receptors. Science 340, 615 (2013).
Wang, C. et al. Structural basis for molecular recognition at serotonin receptors. Science 340, 610 (2013). These studies 84,85 describe crystal structures of 5-HT 1B and 5-HT 2B GPCRs, which provide the first direct structural insights into proximal molecular triggers of biased agonism
Wacker, D. et al. Crystal structure of an LSD-bound human serotonin receptor. Cell 168, 377–389 (2017).
Warne, T., Edwards, P. C., Leslie, A. G. & Tate, C. G. Crystal structures of a stabilized β1-adrenoceptor bound to the biased agonists bucindolol and carvedilol. Structure 20, 841–849 (2012).
Nuber, S. et al. β-Arrestin biosensors reveal a rapid, receptor-dependent activation/deactivation cycle. Nature 531, 661–664 (2016).
Lee, M.-H. et al. The conformational signature of β-arrestin2 predicts its trafficking and signalling functions. Nature 531, 665–668 (2016). These papers 88,89 describe BRET biosensor studies that show evidence for biased agonism arising at the level of the transducer
Lane, J. R., May, L. T., Parton, R. G., Sexton, P. M. & Christopoulos, A. A kinetic view of GPCR allostery and biased agonism. Nat. Chem. Biol. 13, 929–937 (2017).
Klein Herenbrink, C. et al. The role of kinetic context in apparent biased agonism at GPCRs. Nat. Commun. 7, 10842 (2016).
Furness, S. G. et al. Ligand-dependent modulation of G protein conformation alters drug efficacy. Cell 167, 739–749 (2016). This study provides evidence supporting a role for ligand–GPCR interactions in promoting conformational selection at the level of the G protein as an unappreciated molecular mechanism underlying drug efficacy
Gregorio, G. G. et al. Single-molecule analysis of ligand efficacy in β2AR–G-protein activation. Nature 547, 68–73 (2017).
Liang, Y. L. et al. Phase-plate cryo-EM structure of a biased agonist-bound human GLP-1 receptor–Gs complex. Nature 555, 121–125 (2018).
Cai, Y. et al. Purification of family B G protein-coupled receptors using nanodiscs: Application to human glucagon-like peptide-1 receptor. PLoS One 12, e0179568 (2017).
Whorton, M. R. et al. A monomeric G protein-coupled receptor isolated in a high-density lipoprotein particle efficiently activates its G protein. Proc. Natl Acad. Sci. USA 104, 7682–7687 (2007). This key pharmacology study shows that a monomeric class A GPCR can activate and signal through G proteins
Farran, B. An update on the physiological and therapeutic relevance of GPCR oligomers. Pharmacol. Res. 117, 303–327 (2017).
Harikumar, K. G., Ball, A. M., Sexton, P. M. & Miller, L. J. Importance of lipid-exposed residues in transmembrane segment four for family B calcitonin receptor homo-dimerization. Regul. Pept. 164, 113–119 (2010).
Pin, J. P. & Bettler, B. Organization and functions of mGlu and GABAB receptor complexes. Nature 540, 60–68 (2016).
Zhang, C. et al. Structural basis for regulation of human calcium-sensing receptor by magnesium ions and an unexpected tryptophan derivative co-agonist. Sci. Adv. 2, e1600241 (2016).
Geng, Y. et al. Structural mechanism of ligand activation in human calcium-sensing receptor. eLife 5, e13662 (2016).
Nuemket, N. et al. Structural basis for perception of diverse chemical substances by T1r taste receptors. Nat. Commun. 8, 15530 (2017).
Freed, D. M. et al. EGFR ligands differentially stabilize receptor dimers to specify signaling kinetics. Cell 171, 683–695 (2017).
Congreve, M., Oswald, C. & Marshall, F. H. Applying structure-based drug design approaches to allosteric modulators of GPCRs. Trends Pharmacol. Sci. 38, 837–847 (2017).
Roth, B. L., Irwin, J. J. & Shoichet, B. K. Discovery of new GPCR ligands to illuminate new biology. Nat. Chem. Biol. 13, 1143–1151 (2017).
Korczynska, M. et al. Structure-based discovery of selective positive allosteric modulators of antagonists for the M2 muscarinic acetylcholine receptor. Proc. Natl. Acad. Sci. USA 115, E2419–E2428 (2018).
We are grateful to J. Tesmer for assistance with preparation of Supplementary Table 1. This work was funded by the National Health and Medical Research Council of Australia (NHMRC) (Program Grant number APP1055134). D.M.T. and A.G. are Australian Research Council Discovery Early Career Research Award Fellows, P.M.S. is a NHMRC Principal Research Fellow and A.C. is a NHMRC Senior Principal Research Fellow.
Nature thanks A. Jazayeri, R. Lefkowitz and A. Manglik for their contribution to the peer review of this work.
The authors declare no competing interests.
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Thal, D.M., Glukhova, A., Sexton, P.M. et al. Structural insights into G-protein-coupled receptor allostery. Nature 559, 45–53 (2018). https://doi.org/10.1038/s41586-018-0259-z
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