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The molecular basis of subtype selectivity of human kinin G-protein-coupled receptors

Nature Chemical Biology volume 14, pages 284–290 (2018)Cite this article

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Abstract

G-protein-coupled receptors (GPCRs) are the most important signal transducers in higher eukaryotes. Despite considerable progress, the molecular basis of subtype-specific ligand selectivity, especially for peptide receptors, remains unknown. Here, by integrating DNP-enhanced solid-state NMR spectroscopy with advanced molecular modeling and docking, the mechanism of the subtype selectivity of human bradykinin receptors for their peptide agonists has been resolved. The conserved middle segments of the bound peptides show distinct conformations that result in different presentations of their N and C termini toward their receptors. Analysis of the peptide–receptor interfaces reveals that the charged N-terminal residues of the peptides are mainly selected through electrostatic interactions, whereas the C-terminal segments are recognized via both conformations and interactions. The detailed molecular picture obtained by this approach opens a new gateway for exploring the complex conformational and chemical space of peptides and peptide analogs for designing GPCR subtype-selective biochemical tools and drugs.

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Figure 1: Affinities of kinin peptides for their respective human bradykinin receptors, B1R and B2R.
Figure 2: Experimental setup and exemplary spectra of B1R in complex with DAKD.
Figure 3: Backbone structures of DAKD in complex with human B1R in comparison to BK bound to human B2R.
Figure 4: Functional characterization of peptide variants and B1R mutants.
Figure 5: Structural characterization of the B1R–DAKD (green) and B2R–BK (purple) binding pocket.
Figure 6: Representation of key interactions responsible for high affinity binding of DAKD to B1R and of BK to B2R.

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Acknowledgements

We would like to thank T. Mosler and M. Radloff for excellent technical assistance. The German Research Foundation has supported this work through an equipment grant (GL 307/8-1). Funding by DFG project G-NMR and by SFB 807 “Transport and communication across membranes,” the Cluster of Excellence Frankfurt Macromolecular Complexes and the Max Planck Society is acknowledged. The work was also supported by BMRZ through infrastructure support by the State of Hesse. Work in the Meiler laboratory is supported through NIH (R01 GM080403, R01 GM099842 and R01 GM073151) and NSF (CHE 1305874).

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Author notes

  1. Lisa Joedicke

    Present address: UCB Celltech, Slough, Berkshire, UK

  2. Julia Preu

    Present address: Department of Experimental Biomedicine, Rudolf Virchow Center, University of Würzburg, Würzburg, Germany

  3. Lisa Joedicke, Jiafei Mao and Georg Kuenze: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Frankfurt am Main, Germany

    Lisa Joedicke, Christoph Reinhart, Tejaswi Kalavacherla, Julia Preu & Hartmut Michel

  2. Institute of Biophysical Chemistry, Goethe University Frankfurt, Frankfurt am Main, Germany

    Jiafei Mao & Clemens Glaubitz

  3. Centre for Biomolecular Magnetic Resonance, Goethe University Frankfurt, Frankfurt am Main, Germany

    Jiafei Mao, Hendrik R A Jonker, Christian Richter, Harald Schwalbe & Clemens Glaubitz

  4. Center for Structural Biology, Vanderbilt University, Nashville, USA

    Georg Kuenze & Jens Meiler

  5. Institute for Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt am Main, Germany

    Hendrik R A Jonker, Christian Richter & Harald Schwalbe

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Contributions

C.G. and H.M. conceived the project. L.J. expressed and purified B1R and performed radioactive ligand binding assays. J. Mao designed and performed DNP-ssNMR measurements, analyzed data, determined structural models of the kinin peptides and performed the analysis of B1R homolog sequences. G.K. performed molecular modeling and computational docking studies. C.R. and T.K. carried out expression of mutants and additional binding assays. H.R.A.J., C.R. and H.S. provided and analyzed supplementary liquid-state NMR data on nonbound DAKD. J.P. assisted with receptor purification and sample preparation. J. Meiler, H.M., C.G. supervised the overall project. L.J., J. Mao, G.K. and C.G. wrote the manuscript.

Corresponding authors

Correspondence to Jiafei Mao, Hartmut Michel or Clemens Glaubitz.

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Joedicke, L., Mao, J., Kuenze, G. et al. The molecular basis of subtype selectivity of human kinin G-protein-coupled receptors. Nat Chem Biol 14, 284–290 (2018). https://doi.org/10.1038/nchembio.2551

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