Letter | Published:

Structure of the adenosine A2A receptor bound to an engineered G protein

Nature 536, 104–107 (04 August 2016) | Download Citation

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  • An Erratum to this article was published on 14 September 2016

Abstract

G-protein-coupled receptors (GPCRs) are essential components of the signalling network throughout the body. To understand the molecular mechanism of G-protein-mediated signalling, solved structures of receptors in inactive conformations and in the active conformation coupled to a G protein are necessary1,2. Here we present the structure of the adenosine A2A receptor (A2AR) bound to an engineered G protein, mini-Gs, at 3.4 Å resolution. Mini-Gs binds to A2AR through an extensive interface (1,048 Å2) that is similar, but not identical, to the interface between Gs and the β2-adrenergic receptor3. The transition of the receptor from an agonist-bound active-intermediate state4,5 to an active G-protein-bound state is characterized by a 14 Å shift of the cytoplasmic end of transmembrane helix 6 (H6) away from the receptor core, slight changes in the positions of the cytoplasmic ends of H5 and H7 and rotamer changes of the amino acid side chains Arg3.50, Tyr5.58 and Tyr7.53. There are no substantial differences in the extracellular half of the receptor around the ligand binding pocket. The A2AR–mini-Gs structure highlights both the diversity and similarity in G-protein coupling to GPCRs6 and hints at the potential complexity of the molecular basis for G-protein specificity.

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Accessions

Primary accessions

Protein Data Bank

Data deposits

Atomic co-ordinates and structure factors for the A2AR–mini-Gs complex have been submitted to the Protein Data Bank (PDB) under accession code 5G53.

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Acknowledgements

This work was supported by a grant from Heptares Therapeutics Ltd., the ERC (grant EMPSI 339995) and core funding from the Medical Research Council (MC_U105197215 and MC_U105184325). We thank the beamline staff at the European Synchrotron Radiation Facility (beamlines ID23-2, ID30-A3 and ID29) and at Diamond Light Source (beamline I24). We also thank R. Henderson, A. Jazayeri and T. Flock for comments on the manuscript.

Author information

Affiliations

  1. MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK

    • Byron Carpenter
    • , Rony Nehmé
    • , Tony Warne
    • , Andrew G. W. Leslie
    •  & Christopher G. Tate

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Contributions

B.C. performed receptor expression, purification, crystallization, cryo-cooling of the crystals, data collection, data processing and structure refinement. T.W. helped with crystallization, data collection and data processing. R.N. performed the stability assays and pharmacological analyses on A2AR–mini-GS complexes. A.G.W.L. was involved in data processing and structure solution, refinement and analysis. Manuscript preparation was performed by B.C., A.G.W.L. and C.G.T. The overall project management was by C.G.T.

Competing interests

This work was funded in part by Heptares Therapeutics Ltd. C.G.T. is a consultant and member of the Scientific Advisory Board for Heptares Therapeutics Ltd.

Corresponding authors

Correspondence to Byron Carpenter or Christopher G. Tate.

Reviewer Information Nature thanks O. Ernst, K. Jacobson and K.-N. Klotz for their contribution to the peer review of this work.

Extended data

Extended data figures

  1. 1.

    Pharmacological analyses of A2AR–mini-Gs complexes.

  2. 2.

    Thermostability of detergent-solubilized 3H-NECA-bound A2AR in the presence or absence of mini-Gs.

  3. 3.

    Omit maps for NECA and GDP.

  4. 4.

    Electron density for the interface region of the A2AR–mini-Gs complex.

  5. 5.

    Alignment of mini-Gs with GNAS2.

  6. 6.

    Alignment of β2AR and A2AR amino acid sequences.

  7. 7.

    A conserved hydrophobic binding pocket at the receptor–Gαs interface.

  8. 8.

    Comparison between receptor-bound mini-Gs and Gαs.

  9. 9.

    Comparison of the NECA binding site in the active-intermediate state compared to the mini-Gs-bound state.

Extended data tables

  1. 1.

    Data collection and refinement statistics

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains a Supplementary Discussion.

Excel files

  1. 1.

    Supplementary Table

    This file contains Supplementary Table 1.

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DOI

https://doi.org/10.1038/nature18966

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