CC chemokine receptor 2 (CCR2) is one of 19 members of the chemokine receptor subfamily of human class A G-protein-coupled receptors. CCR2 is expressed on monocytes, immature dendritic cells, and T-cell subpopulations, and mediates their migration towards endogenous CC chemokine ligands such as CCL2 (ref. 1). CCR2 and its ligands are implicated in numerous inflammatory and neurodegenerative diseases2 including atherosclerosis, multiple sclerosis, asthma, neuropathic pain, and diabetic nephropathy, as well as cancer3. These disease associations have motivated numerous preclinical studies and clinical trials4 (see http://www.clinicaltrials.gov) in search of therapies that target the CCR2–chemokine axis. To aid drug discovery efforts5, here we solve a structure of CCR2 in a ternary complex with an orthosteric (BMS-681 (ref. 6)) and allosteric (CCR2-RA-[R]7) antagonist. BMS-681 inhibits chemokine binding by occupying the orthosteric pocket of the receptor in a previously unseen binding mode. CCR2-RA-[R] binds in a novel, highly druggable pocket that is the most intracellular allosteric site observed in class A G-protein-coupled receptors so far; this site spatially overlaps the G-protein-binding site in homologous receptors. CCR2-RA-[R] inhibits CCR2 non-competitively by blocking activation-associated conformational changes and formation of the G-protein-binding interface. The conformational signature of the conserved microswitch residues observed in double-antagonist-bound CCR2 resembles the most inactive G-protein-coupled receptor structures solved so far. Like other protein–protein interactions, receptor–chemokine complexes are considered challenging therapeutic targets for small molecules, and the present structure suggests diverse pocket epitopes that can be exploited to overcome obstacles in drug design.

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  1. 1.

    et al. Pharmacological modulation of chemokine receptor function. Br. J. Pharmacol. 165, 1617–1643 (2012)

  2. 2.

    , & CCL2-CCR2 signaling in disease pathogenesis. Endocr. Metab. Immune Disord. Drug Targets 15, 105–118 (2015)

  3. 3.

    , , & Targeting the CCL2-CCR2 signaling axis in cancer metastasis. Oncotarget 7, 28697–28710 (2016)

  4. 4.

    , & “Chemokine receptors as therapeutic targets: why aren’t there more drugs?”. Eur. J. Pharmacol. 746, 363–367 (2015)

  5. 5.

    , , & Structures of G protein-coupled receptors reveal new opportunities for drug discovery. Drug Discov. Today 20, 1355–1364 (2015)

  6. 6.

    et al. Discovery of a potent and orally bioavailable dual antagonist of CC chemokine receptors 2 and 5. ACS Med. Chem. Lett. 6, 439–444 (2015)

  7. 7.

    et al. Novel, acidic CCR2 receptor antagonists: lead optimization. Lett. Drug Des. Discov. 4, 263–271 (2007)

  8. 8.

    & Crystallizing membrane proteins using lipidic mesophases. Nature Protocols 4, 706–731 (2009)

  9. 9.

    et al. Structures of the CXCR4 chemokine GPCR with small-molecule and cyclic peptide antagonists. Science 330, 1066–1071 (2010)

  10. 10.

    et al. Structure of the CCR5 chemokine receptor-HIV entry inhibitor maraviroc complex. Science 341, 1387–1390 (2013)

  11. 11.

    et al. Structural biology. Crystal structure of the chemokine receptor CXCR4 in complex with a viral chemokine. Science 347, 1117–1122 (2015)

  12. 12.

    et al. Structural basis for chemokine recognition and activation of a viral G protein-coupled receptor. Science 347, 1113–1117 (2015)

  13. 13.

    & The amino-terminal domain of CCR2 is both necessary and sufficient for high affinity binding of monocyte chemoattractant protein 1. Receptor activation by a pseudo-tethered ligand. J. Biol. Chem. 272, 23186–23190 (1997)

  14. 14.

    et al. Structure of the human P2Y12 receptor in complex with an antithrombotic drug. Nature 509, 115–118 (2014)

  15. 15.

    et al. Specific chemical and structural damage to proteins produced by synchrotron radiation. Proc. Natl Acad. Sci. USA 97, 623–628 (2000)

  16. 16.

    et al. Discovery of disubstituted cyclohexanes as a new class of CC chemokine receptor 2 antagonists. J. Med. Chem. 51, 721–724 (2008)

  17. 17.

    et al. CCR2: characterization of the antagonist binding site from a combined receptor modeling/mutagenesis approach. J. Med. Chem. 46, 4070–4086 (2003)

  18. 18.

    et al. Elucidation of binding sites of dual antagonists in the human chemokine receptors CCR2 and CCR5. Mol. Pharmacol. 75, 1325–1336 (2009)

  19. 19.

    et al. Synthesis and evaluation of cis-3,4-disubstituted piperidines as potent CC chemokine receptor 2 (CCR2) antagonists. Bioorg. Med. Chem. Lett. 18, 5063–5065 (2008)

  20. 20.

    et al. Discovery and mapping of an intracellular antagonist binding site at the chemokine receptor CCR2. Mol. Pharmacol. 86, 358–368 (2014)

  21. 21.

    , & The high-resolution structure of activated opsin reveals a conserved solvent network in the transmembrane region essential for activation. Structure 23, 2358–2364 (2015)

  22. 22.

    et al. Crystal structure of the β2 adrenergic receptor–Gs protein complex. Nature 477, 549–555 (2011)

  23. 23.

    et al. Multiple binding sites for small-molecule antagonists at the CC chemokine receptor 2. Mol. Pharmacol. 84, 551–561 (2013)

  24. 24.

    et al. G-protein-coupled receptor inactivation by an allosteric inverse-agonist antibody. Nature 482, 237–240 (2012)

  25. 25.

    et al. Allosteric nanobodies reveal the dynamic range and diverse mechanisms of G-protein-coupled receptor activation. Nature 535, 448–452 (2016)

  26. 26.

    , & Structure-function of the G protein-coupled receptor superfamily. Annu. Rev. Pharmacol. Toxicol. 53, 531–556 (2013)

  27. 27.

    et al. Structural insights into the dynamic process of β2-adrenergic receptor signaling. Cell 161, 1101–1111 (2015)

  28. 28.

    et al. Crystal structure of rhodopsin: a G protein-coupled receptor. Science 289, 739–745 (2000)

  29. 29.

    et al. Crystal structure of the human β2 adrenergic G-protein-coupled receptor. Nature 450, 383–387 (2007)

  30. 30.

    & Chemokine receptor antagonists. J. Med. Chem. 55, 9363–9392 (2012)

  31. 31.

    , & An intracellular allosteric site for a specific class of antagonists of the CC chemokine G protein-coupled receptors CCR4 and CCR5. Mol. Pharmacol. 73, 855–867 (2008)

  32. 32.

    et al. Identification of a putative intracellular allosteric antagonist binding-site in the CXC chemokine receptors 1 and 2. Mol. Pharmacol. 74, 1193–1202 (2008)

  33. 33.

    , , , & Microscale fluorescent thermal stability assay for membrane proteins. Structure 16, 351–359 (2008)

  34. 34.

    et al. Rastering strategy for screening and centring of microcrystal samples of human membrane proteins with a sub-10 μm size X-ray synchrotron beam. J. R. Soc. Interface 6 (Suppl. 5), S587–S597 (2009)

  35. 35.

    Xds. Acta Crystallogr. D 66, 125–132 (2010)

  36. 36.

    et al. Overview of the CCP4 suite and current developments. Acta Crystallogr. D 67, 235–242 (2011)

  37. 37.

    et al. Phaser crystallographic software. J. Appl. Crystallogr. 40, 658–674 (2007)

  38. 38.

    et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr. D 66, 213–221 (2010)

  39. 39.

    , , & Features and development of Coot. Acta Crystallogr. D 66, 486–501 (2010)

  40. 40.

    et al. Discovery of potent, orally bioavailable small-molecule inhibitors of the human CCR2 receptor. ChemMedChem 3, 660–669 (2008)

  41. 41.

    et al. Discovery and pharmacological characterization of a novel rodent-active CCR2 antagonist, INCB3344. J. Immunol. 175, 5370–5378 (2005)

  42. 42.

    et al. MolProbity: all-atom structure validation for macromolecular crystallography. Acta Crystallogr. D 66, 12–21 (2010)

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We thank A. Ishchenko and H. Zhang for help with X-ray data collection, C. Wang and H. X. Wu for suggestions on construct design, F. Li for help with data processing, and M. Galella for assistance with BMS compound data and statistics. We thank C. Ogata, R. Sanishvili, N. Venugopalan, M. Becker, and S. Corcoran at beamline 23ID at GM/CA CAT Advanced Photon Source. Funding for this research was provided by National Institutes of Health grants R01 GM071872, U54 GM094618, R01 AI118985, R21 AI121918, and R21 AI122211. GM/CA@APS has been funded in whole or in part with federal funds from the National Cancer Institute (ACB-12002) and the National Institute of General Medical Sciences (AGM-12006). This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under contract number DE-AC02-06CH11357.

Author information


  1. Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA

    • Yi Zheng
    • , Ling Qin
    • , Martin Gustavsson
    • , Chunxia Zhao
    • , Ruben Abagyan
    • , Irina Kufareva
    •  & Tracy M. Handel
  2. Division of Medicinal Chemistry, Leiden Academic Centre for Drug Research (LACDR), Leiden University, Leiden 2333 CC, The Netherlands

    • Natalia V. Ortiz Zacarías
    • , Henk de Vries
    • , Adriaan P. IJzerman
    •  & Laura H. Heitman
  3. Bridge Institute, Departments of Chemistry and Physics & Astronomy, University of Southern California, Los Angeles, California 90089, USA

    • Gye Won Han
    •  & Vadim Cherezov
  4. Bristol-Myers Squibb Company, Princeton, New Jersey 08543, USA

    • Marta Dabros
    • , Robert J. Cherney
    • , Percy Carter
    •  & Andrew Tebben
  5. Vertex Pharmaceuticals Inc., 11010 Torreyana Road, San Diego, California 92121, USA

    • Dean Stamos
  6. The Bridge Institute, Departments of Biological Sciences and Chemistry, University of Southern California, Los Angeles, California 90089, USA

    • Raymond C. Stevens


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I.K. and T.M.H. designed the study and coordinated all experiments. Y.Z. designed and engineered protein constructs, performed crystallization experiments, collected the diffraction data, and determined the structure. L.Q., M.G., and C.Z. assisted with protein engineering and crystallization. G.W.H. assisted with structure determination and refinement. A.P.I. and L.H.H. designed, and N.V.O.Z. and H.d.V. performed, equilibrium and kinetics binding experiments. I.K. performed computational and bioinformatics analyses. R.J.C., P.C., and A.T. synthesized, characterized, and crystallized the BMS compound analogues. M.D. assisted with compound crystallization. D.S. assisted with the allosteric compound characterization. R.A. assisted with structure analysis. V.C. and R.C.S. assisted with crystallization. Y.Z., N.V.O.Z., A.P.I., L.H.H., I.K., and T.M.H. wrote the paper.

Competing interests

R.A. has an equity interest in Molsoft, LLC. The terms of this arrangement have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. R.C., P.C., and A.T. are employees of Bristol-Myers Squibb Company. D.S. is an employee of Vertex Pharmaceuticals, Inc.

Corresponding authors

Correspondence to Laura H. Heitman or Irina Kufareva or Tracy M. Handel.

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