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Dual agonist occupancy of AT1-R–α2C-AR heterodimers results in atypical Gs-PKA signaling

Nature Chemical Biology volume 11, pages 271–279 (2015)Cite this article

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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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Figure 1: Characterization of the heterodimerization between AT1-R and α2C-AR in HEK293T cells.
Figure 2: Characterization of individual agonist effects on the AT1-R–α2C-AR heterodimer.
Figure 3: CODA-RET characterization of β-arrestin 2 and G-protein recruitment to heteromeric α2C and AT1 receptors.
Figure 4: The nature of ligands dictates different β-arrestin 2 trafficking pathways activated by the AT1-R–α2C-AR heterodimer.
Figure 5: AngII-NE costimulation of the AT1-R–α2C-AR heterodimer generates new Gs-cAMP-PKA signaling.
Figure 6: NE potentiates AngII-induced increase in NE release in neurons or RSNA in mice through a PKA pathway.

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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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Author notes

  1. Ségolène Galandrin, Cédric Boularan, Heinrich J Matthies and Fabien Despas: These authors contributed equally to this work.

Authors and Affiliations

  1. Institut des Maladies Métaboliques et Cardiovasculaires, Institut National de la Santé et de la Recherche Médicale, U1048, Université Toulouse III Paul Sabatier, Toulouse, France

    Morgane Bellot, Ségolène Galandrin, Cédric Boularan, Fabien Despas, Colette Denis, Véronique Pons, Marie-Hélène Seguelas, Atul Pathak, Jean-Michel Sénard & Céline Galés

  2. Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Heinrich J Matthies & Aurelio Galli

  3. Neuroscience Program in Substance Abuse, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Heinrich J Matthies & Aurelio Galli

  4. Service de Pharmacologie Clinique, Centre Hospitalier Universitaire de Toulouse, Faculté de Médecine, Toulouse, France

    Fabien Despas, Atul Pathak & Jean-Michel Sénard

  5. Center for Molecular Recognition and Departments of Psychiatry and Pharmacology, Columbia University, New York, New York, USA

    Jonathan Javitch

  6. College of Physicians and Surgeons, Columbia University, New York, New York, USA

    Jonathan Javitch

  7. Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York, USA

    Jonathan Javitch

  8. Institut de Pharmacologie et de Biologie Structurale (IPBS), CNRS, UMR 508, Université Toulouse III Paul Sabatier, Toulouse, France

    Serge Mazères

  9. Department of Clinical Biochemistry, Glostrup Hospital, Glostrup, Denmark

    Samra Joke Sanni

  10. Diabetes Biology and Metabolism, Novo Nordisk, Måløv, Denmark

    Samra Joke Sanni & Jakob L Hansen

  11. Vanderbilt Brain Institute, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Aurelio Galli

  12. Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Aurelio Galli

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Contributions

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.

Corresponding author

Correspondence to Céline Galés.

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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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