Abstract
Hypersecretion of norepinephrine (NE) and angiotensin II (AngII) is a hallmark of major prevalent cardiovascular diseases that contribute to cardiac pathophysiology and morbidity. Herein, we explore whether heterodimerization of presynaptic AngII AT1 receptor (AT1-R) and NE α2C-adrenergic receptor (α2C-AR) could underlie their functional cross-talk to control NE secretion. Multiple bioluminescence resonance energy transfer and protein complementation assays allowed us to accurately probe the structures and functions of the α2C-AR–AT1-R dimer promoted by ligand binding to individual protomers. We found that dual agonist occupancy resulted in a conformation of the heterodimer different from that induced by active individual protomers and triggered atypical Gs-cAMP–PKA signaling. This specific pharmacological signaling unit was identified in vivo to promote not only NE hypersecretion in sympathetic neurons but also sympathetic hyperactivity in mice. Thus, we uncovered a new process by which GPCR heterodimerization creates an original functional pharmacological entity and that could constitute a promising new target in cardiovascular therapeutics.
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Acknowledgements
M.B. was supported by a doctoral fellow fellowship from the Fondation pour la Recherche Médicale. J.-M.S. and C.G. are supported by the Institut National de la Santé et de la Recherche Médicale and by grants from La Société Française d'Hypertension Artérielle and the Fondation Bettencourt Schueller. H.J.M. was supported by a US National Institutes of Health grant (grant number DA038058-01).
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M.B. designed and performed most of the experiments, analyzed and interpreted data and helped to write the manuscript. S.G. and C.B. helped in the anisotropy experiments and performed β-arrestin and cAMP experiments, analyzed data and helped to write the manuscript. H.J.M. performed NE-release experiments in primary cultures of sympathetic neurons and analyzed the data. F.D. designed, performed and analyzed mouse microneurography experiments. C.D. helped in the binding studies, the design of most experiments and the interpretation of the data. J.J. helped with construction of the Venus and Luciferase fusion BRET probes, the interpretation of the data and the editing of the manuscript. S.M. performed anisotropy experiments and analyzed the data. V.P. and M.-H.S. established some plasmid constructs. S.J.S. and J.L.H. performed some binding studies and analyzed and interpreted data. A.P. assisted in data processing and analysis and with manuscript preparation. A.G. supervised the neuron experiments and wrote the manuscript. J.-M.S. supervised some aspects of the project, analyzed the data and wrote the manuscript. C.G. conceived and supervised the project, performed data analysis and wrote the manuscript.
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J.L.H. is an employee of Novo Nordisk and is a shareholder of Novo Nordisk.
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Bellot, M., Galandrin, S., Boularan, C. et al. Dual agonist occupancy of AT1-R–α2C-AR heterodimers results in atypical Gs-PKA signaling. Nat Chem Biol 11, 271–279 (2015). https://doi.org/10.1038/nchembio.1766
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DOI: https://doi.org/10.1038/nchembio.1766
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