Abstract
We present a new concept of partial agonism at G protein–coupled receptors. We demonstrate the coexistence of two functionally distinct populations of the muscarinic M2 receptor stabilized by one dynamic ligand, which binds in two opposite orientations. The ratio of orientations determines the cellular response. Our concept allows predicting and virtually titrating ligand efficacy, which opens unprecedented opportunities for the design of drugs with graded activation of the biological system.
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References
Pierce, K.L., Premont, R.T. & Lefkowitz, R.J. Nat. Rev. Mol. Cell Biol. 3, 639–650 (2002).
Venkatakrishnan, A.J. et al. Nature 494, 185–194 (2013).
Kenakin, T. Nat. Rev. Drug Discov. 1, 103–110 (2002).
Kenakin, T. & Christopoulos, A. Nat. Rev. Drug Discov. 12, 205–216 (2013).
Rosenbaum, D.M., Rasmussen, S.G.F. & Kobilka, B.K. Nature 459, 356–363 (2009).
Warne, T. et al. Nature 469, 241–244 (2011).
Swaminath, G. et al. J. Biol. Chem. 280, 22165–22171 (2005).
Yao, X. et al. Nat. Chem. Biol. 2, 417–422 (2006).
Kofuku, Y. et al. Nat. Commun. 3, 1045 (2012).
Nikolaev, V.O., Hoffmann, C., Bünemann, M., Lohse, M.J. & Vilardaga, J.-P. J. Biol. Chem. 281, 24506–24511 (2006).
Zürn, A. et al. Mol. Pharmacol. 75, 534–541 (2009).
Kenakin, T. & Miller, L.J. Pharmacol. Rev. 62, 265–304 (2010).
Nygaard, R. et al. Cell 152, 532–542 (2013).
Black, J.W. & Leff, P. Proc. R. Soc. Lond. B Biol. Sci. 220, 141–162 (1983).
Bruning, J.B. et al. Nat. Chem. Biol. 6, 837–843 (2010).
Overington, J.P., Al-Lazikani, B. & Hopkins, A.L. Nat. Rev. Drug Discov. 5, 993–996 (2006).
Antony, J. et al. FASEB J. 23, 442–450 (2009).
Bock, A. et al. Nat. Commun. 3, 1044 (2012).
Prilla, S., Schrobang, J., Ellis, J., Höltje, H.-D. & Mohr, K. Mol. Pharmacol. 70, 181–193 (2006).
Schröder, R. et al. Nat. Biotechnol. 28, 943–949 (2010).
Schröder, R. et al. Nat. Protoc. 6, 1748–1760 (2011).
Kloeckner, J., Schmitz, J. & Holzgrabe, U. Tetrahedr. Lett. 51, 3470–3472 (2010).
Dallanoce, C. et al. Bioorg. Med. Chem. 7, 1539–1547 (1999).
Disingrini, T. et al. J. Med. Chem. 49, 366–372 (2006).
Schrage, R. et al. Br. J. Pharmacol. 169, 357–370 (2013).
Hoffmann, C. et al. Nat. Methods 2, 171–176 (2005).
Jäger, D. et al. J. Biol. Chem. 282, 34968–34976 (2007).
Ehlert, F.J. Mol. Pharmacol. 33, 187–194 (1988).
May, L.T., Leach, K., Sexton, P.M. & Christopoulos, A. Annu. Rev. Pharmacol. Toxicol. 47, 1–51 (2007).
Ehlert, F.J., Griffin, M.T., Sawyer, G.W. & Bailon, R. J. Pharmacol. Exp. Ther. 289, 981–992 (1999).
Griffin, M.T., Figueroa, K.W., Liller, S. & Ehlert, F.J. J. Pharmacol. Exp. Ther. 321, 1193–1207 (2007).
Acknowledgements
We thank M. Kepe for excellent technical assistance and Corning Inc. for their support on the Epic system. A.B. is a member of the graduate school Theoretical and Experimental Medicine at the University of Bonn. This work was funded by the Deutsche Forschungsgemeinschaft (DFG) by grants to K.M. (MO 821/2-1), U.H. (HO 1368/12-1), E.K. (KO 1583/3-1) and C.H. (SFB487 TPA1). B.C. is funded by the North-Rhine-Westphalia International Graduate Research School BIOTECH-PHARMA at the University of Bonn.
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A.B. conceived the project, developed the mathematical framework for dynamic ligands, conducted all of the binding experiments in live CHO-M2 and CHO-M2W422A cells and all of the [35S]GTPγS binding experiments and DMR assays and supervised experiments related to binding to membranes of CHO-hM2 cells and to characterization of the allosteric fragments. B.C. characterized the allosteric fragments in binding and functional assays and conducted binding assays of the X-6-phth series in membranes of CHO-hM2 cells. F.K. conducted binding assay of the X-6-naph series in membranes of CHO-hM2 cells. R.M. synthesized and characterized 8-naph. J.B. conducted FRET experiments. M.M. synthesized and characterized isox-8-naph. C.D. provided isox and isox-6-phth. D.K. conducted all of the [35S]GTPγS binding experiments to membranes of CHO-M2W422A cells. C.H. planned and supervised FRET experiments. U.H. planned and supervised chemical syntheses. C.T., M.D.A., C.H. and U.H. contributed to discussions. E.K. provided essential ideas. A.B. and K.M. made figures and wrote the manuscript. K.M. supervised the overall research.
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Supplementary information
Supplementary Text and Figures
Supplementary Results, Supplementary Figures 1–11 and Supplementary Tables 1–3. (PDF 1187 kb)
Supplementary Note 1
The binding model for dynamic ligands (PDF 428 kb)
Supplementary Note 2
Chemical syntheses and characterization (PDF 137 kb)
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Bock, A., Chirinda, B., Krebs, F. et al. Dynamic ligand binding dictates partial agonism at a G protein–coupled receptor. Nat Chem Biol 10, 18–20 (2014). https://doi.org/10.1038/nchembio.1384
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DOI: https://doi.org/10.1038/nchembio.1384
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