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Crystal structure of oligomeric β1-adrenergic G protein–coupled receptors in ligand-free basal state

Nature Structural & Molecular Biology volume 20, pages 419–425 (2013)Cite this article

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Abstract

G protein–coupled receptors (GPCRs) mediate transmembrane signaling. Before ligand binding, GPCRs exist in a basal state. Crystal structures of several GPCRs bound with antagonists or agonists have been solved. However, the crystal structure of the ligand-free basal state of a GPCR, the starting point of GPCR activation and function, had not yet been determined. Here we report the X-ray crystal structure of the ligand-free basal state of a GPCR in a lipid membrane–like environment. Oligomeric turkey β1-adrenergic receptors display two dimer interfaces. One interface involves the transmembrane domain (TM) 1, TM2, the C-terminal H8 and extracellular loop 1. The other interface engages residues from TM4, TM5, intracellular loop 2 and extracellular loop 2. Structural comparisons show that this ligand-free state is in an inactive conformation. This provides the structural basis of GPCR dimerization and oligomerization.

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Figure 1: Structure of the ligand-free basal-state β1-AR.
Figure 2: Dimer interface 1 of β1-AR oligomers.
Figure 3: Dimer interface 2 of β1-AR oligomers.
Figure 4: The ligand-free basal state of β1-AR in an inactive conformation and with a contracted ligand-binding pocket.
Figure 5: Docking of Gs onto the β1-AR dimer.
Figure 6: Comparison of the β1-AR oligomer with the μ-opioid receptor oligomer.

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Acknowledgements

We thank O. Andersen, O. Boudker, J. McCoy, C. Steegborn, Y. Sun, G. Verdon and W. Xu for advice, discussions and help. We thank E. Ross (University of Texas Southwestern Medical Center, Dallas, Texas, USA) for the turkey β1-AR plasmid, E. Gouaux (Vollum Institute, Portland, Oregon, USA) and O. Boudker (Weill Cornell Medical College, New York, New York, USA) for the pCGFP-EU plasmid and Cornell's chemistry core facility for the synthesis of alprenolol-NH2. We thank I. Kourinov at the Advanced Photon Source beamline 24-ID-E and J. Jakoncic at the Brookhaven National Laboratory NSLS beamlines X6A and X25 for their assistance with X-ray data collection. We are grateful to O. Boudker, H. Weinstein and members of our laboratory for critically reading the manuscript. This work was supported by US National Institutes of Health grant HL 91525 (X.-Y.H.).

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  1. Department of Physiology and Biophysics, Cornell University Weill Medical College, New York, New York, USA

    Jianyun Huang, Shuai Chen, J Jillian Zhang & Xin-Yun Huang

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Contributions

J.H. performed the protein purification, set up crystallization trials, grew crystals for data collection and participated in data collection. S.C. processed the diffraction data, and solved and refined the structures. J.J.Z. participated in project strategy and manuscript preparation. X.-Y.H. was responsible for the overall project strategy and management and participated in data collection and wrote the manuscript.

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Correspondence to Xin-Yun Huang.

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Huang, J., Chen, S., Zhang, J. et al. Crystal structure of oligomeric β1-adrenergic G protein–coupled receptors in ligand-free basal state. Nat Struct Mol Biol 20, 419–425 (2013). https://doi.org/10.1038/nsmb.2504

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