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Structure of class B GPCR corticotropin-releasing factor receptor 1

Nature volume 499, pages 438–443 (2013)Cite this article

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Abstract

Structural analysis of class B G-protein-coupled receptors (GPCRs), cell-surface proteins that respond to peptide hormones, has been restricted to the amino-terminal extracellular domain, thus providing little understanding of the membrane-spanning signal transduction domain. The corticotropin-releasing factor receptor type 1 is a class B receptor which mediates the response to stress and has been considered a drug target for depression and anxiety. Here we report the crystal structure of the transmembrane domain of the human corticotropin-releasing factor receptor type 1 in complex with the small-molecule antagonist CP-376395. The structure provides detailed insight into the architecture of class B receptors. Atomic details of the interactions of the receptor with the non-peptide ligand that binds deep within the receptor are described. This structure provides a model for all class B GPCRs and may aid in the design of new small-molecule drugs for diseases of brain and metabolism.

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Figure 1: Schematic representation of the CRF1R structure.
Figure 2: Overall structure.
Figure 3: Comparison of the antagonist-bound structures of CRF1R and D3R.
Figure 4: Conserved sequence motifs.
Figure 5: Antagonist-binding site.

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Protein Data Bank

Data deposits

Co-ordinates and structure factors have been deposited in the Protein Data Bank under the accession code 4K5Y.

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Acknowledgements

We thank G. Evans, R. Owen and D. Axford for their support. We are grateful to A. Leslie, R. Read and A. McCoy for advice on data collection and structure determination. We thank R. Henderson, A. Leslie, C. Tate as well as F. Blaney, B. Tehan, R. Miller, F. Deflorian and other colleagues for suggestions and comments.

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Authors and Affiliations

  1. Heptares Therapeutics Ltd, BioPark, Broadwater Road, Welwyn Garden City, AL7 3AX, UK,

    Kaspar Hollenstein, James Kean, Andrea Bortolato, Robert K. Y. Cheng, Andrew S. Doré, Ali Jazayeri, Robert M. Cooke, Malcolm Weir & Fiona H. Marshall

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Contributions

J.K. carried out the conformational thermostabilization and pharmacological characterization of the constructs and determined the stability of the StaR in a panel of reagents/additives to aid purification and crystallization. A.J. constructed the T4 lysozyme fusion proteins and identified the usefulness of the fusion into intracellular loop 2. K.H. designed and characterized truncation constructs, designed the final T4 lysozyme fusion construct, established procedures for, and carried out expression and purification, established the platform/protocols for and carried out lipidic cubic phase (LCP) crystallization, collected and processed X-ray diffraction data, solved and refined the structure. R.K.Y.C. performed expression and purification, optimized purification and grew crystals in LCP for data collection of the final construct, collected X-ray diffraction data and solved and refined the structure. A.S.D. was involved in construct design, established the platform/protocols for, and carried out LCP crystallization, collected and processed X-ray diffraction data and solved and refined the structure. Computational analysis of the structure and modelling was carried out by A.B. Project management was carried out by A.J., R.M.C., M.W. and F.H.M. The manuscript was prepared by K.H., R.K.Y.C., A.B., J.K., A.J. and F.H.M.

Corresponding author

Correspondence to Fiona H. Marshall.

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Competing interests

The authors are employees and shareholders of Heptares Therapeutics Ltd, a GPCR drug discovery company using structure-based drug design techniques.

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Hollenstein, K., Kean, J., Bortolato, A. et al. Structure of class B GPCR corticotropin-releasing factor receptor 1. Nature 499, 438–443 (2013). https://doi.org/10.1038/nature12357

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