Abstract
Differential targeting of heterotrimeric G protein versus β-arrestin signaling are emerging concepts in G protein–coupled receptor (GPCR) research and drug discovery, and biased engagement by GPCR ligands of either β-arrestin or G protein pathways has been disclosed. Herein we report on a new mechanism of ligand bias to titrate the signaling specificity of a cell-surface GPCR. Using a combination of biomolecular and virtual screening, we identified the small-molecule modulator Gue1654, which inhibits Gβγ but not Gα signaling triggered upon activation of Gαi-βγ by the chemoattractant receptor OXE-R in both recombinant and human primary cells. Gue1654 does not interfere nonspecifically with signaling directly at or downstream of Gβγ. This hitherto unappreciated mechanism of ligand bias at a GPCR highlights both a new paradigm for functional selectivity and a potentially new strategy to develop pathway-specific therapeutics.
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Acknowledgements
We thank U. Rick and M. Vasmer-Ehses for excellent technical assistance and Corning Inc. for their support on the Epic system. C.D.W. acknowledges E. Days and Y. Du for valuable technical assistance with cell culture and transient transfections. All thallium flux assays were performed in the Vanderbilt Institute of Chemical Biology High-Throughput Screening Facility. This work was funded by a grant from the Austrian Science Funds FWF (P-22521 to A.H.), a grant from the Spanish Ministerio de Ciencia e Innovación (MICINN, SAF2010-22198-C01-02 to L. Pardo), a L'Oreal Fellowship to P.L. and a fellowship of the German Research Foundation (Graduate College 804) to P.A.O.
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S.B. designed and performed experiments and provided important ideas. L. Peters, P.A.O., A.B., V.K., C.D.W., R.S., P.L., J.G. and S.H. designed and performed experiments. A.G. and L. Pardo created the receptor model, performed the virtual screening and contributed to discussion. R.T. and T.U. established synthesis of 5-oxo-ETE, edited the manuscript and contributed to discussion. C.D.W., L. Pardo, K.M., M.G. and A.H. designed research, contributed to discussion and edited the manuscript. E.K. designed research and wrote the manuscript.
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Blättermann, S., Peters, L., Ottersbach, P. et al. A biased ligand for OXE-R uncouples Gα and Gβγ signaling within a heterotrimer. Nat Chem Biol 8, 631–638 (2012). https://doi.org/10.1038/nchembio.962
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DOI: https://doi.org/10.1038/nchembio.962
