Allosteric ligands control the activation of a class C GPCR heterodimer by acting at the transmembrane interface

  1. Lei Liu
  2. Zhiran Fan
  3. Xavier Rovira
  4. Li Xue
  5. Salomé Roux
  6. Isabelle Brabet
  7. Mingxia Xin
  8. Jean-Philippe Pin  Is a corresponding author
  9. Jianfeng Liu  Is a corresponding author
  10. Philippe Rondard  Is a corresponding author
  1. Huazhong University of Science and Technology, China
  2. Spanish National Research Council, Spain
  3. University of Montpellier, CNRS, INSERM, France

Abstract

G protein-coupled receptors (GPCRs) are among the most promising drug targets. They often form homo- and heterodimers with allosteric cross-talk between receptor entities, which contributes to fine tuning of transmembrane signaling. Specifically controlling the activity of GPCR dimers with ligands is a good approach to clarify their physiological roles and to validate them as drug targets. Here, we examined the mode of action of positive allosteric modulators (PAMs) that bind at the interface of the transmembrane domains of the heterodimeric GABAB receptor. Our site-directed mutagenesis results show that mutations of this interface impact the function of the three PAM tested. The data support the inference that they act at the active interface between both transmembrane domains, the binding site involving residues of the TM6s of the GABAB1 and the GABAB2 subunit. Importantly, the agonist activity of these PAMs involves a key region in the central core of the GABAB2 transmembrane domain, which also controls the constitutive activity of the GABAB receptor. This region corresponds to the sodium ion binding site in class A GPCRs that controls the basal state of the receptors. Overall, these data reveal the possibility of developing allosteric compounds able to specifically modulate the activity of GPCR homo- and heterodimers by acting at their transmembrane interface.

Data availability

Figure 2- Source Data 1 contain the numerical data used to generate the figures;Figure 3 - Source Data 1 contain the numerical data used to generate the figures;Figure 4 - Source Data 1 contain the numerical data used to generate the figures;Figure 5 - Source Data 1 contain the numerical data used to generate the figures.

Article and author information

Author details

  1. Lei Liu

    Cellular Signaling Laboratory, International Research Center for Sensory Biology and Technology of MOST, Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9824-9570
  2. Zhiran Fan

    Cellular Signaling Laboratory, International Research Center for Sensory Biology and Technology of MOST, Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9543-1211
  3. Xavier Rovira

    MCS, Laboratory of Medicinal Chemistry, Institute for Advanced Chemistry of Catalonia (IQAC-CSIC), Spanish National Research Council, Barcelona, Spain
    Competing interests
    The authors declare that no competing interests exist.
  4. Li Xue

    University of Montpellier, CNRS, INSERM, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
  5. Salomé Roux

    University of Montpellier, CNRS, INSERM, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6106-4863
  6. Isabelle Brabet

    University of Montpellier, CNRS, INSERM, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
  7. Mingxia Xin

    Cellular Signaling Laboratory, International Research Center for Sensory Biology and Technology of MOST, Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  8. Jean-Philippe Pin

    University of Montpellier, CNRS, INSERM, Montpellier, France
    For correspondence
    jean-philippe.pin@igf.cnrs.fr
    Competing interests
    The authors declare that no competing interests exist.
  9. Jianfeng Liu

    Cellular Signaling Laboratory, International Research Center for Sensory Biology and Technology of MOST, Huazhong University of Science and Technology, Wuhan, China
    For correspondence
    jfliu@mail.hust.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0284-8377
  10. Philippe Rondard

    University of Montpellier, CNRS, INSERM, Montpellier, France
    For correspondence
    philippe.rondard@igf.cnrs.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1134-2738

Funding

Ministry of Science and Technology of the People's Republic of China (2018YFA0507003)

  • Jianfeng Liu

Agence Nationale de la Recherche (ANR-09-PIRI-0011)

  • Philippe Rondard

Fondation pour la recherche médicale FRM (FRM team: DEQ20170326522)

  • Jean-Philippe Pin

Spanish Ministry of Economy (SAF2015-74132-JIN)

  • Xavier Rovira

National Natural Science Foundation of China (81720108031)

  • Jianfeng Liu

National Natural Science Foundation of China (81872945)

  • Jianfeng Liu

National Natural Science Foundation of China (31721002)

  • Jianfeng Liu

National Natural Science Foundation of China (31420103909)

  • Jianfeng Liu

Ministry of Education of the People's Republic of China (B08029)

  • Jianfeng Liu

Centre National de la Recherche Scientifique (PICS n{degree sign}07030)

  • Philippe Rondard

Centre National de la Recherche Scientifique (PRC n{degree sign}1403)

  • Philippe Rondard

Institut National de la Santé et de la Recherche Médicale (IRP Brain Signal)

  • Philippe Rondard

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Andrew C Kruse, Harvard Medical School, United States

Version history

  1. Received: May 9, 2021
  2. Accepted: December 2, 2021
  3. Accepted Manuscript published: December 6, 2021 (version 1)
  4. Accepted Manuscript updated: December 10, 2021 (version 2)
  5. Version of Record published: December 23, 2021 (version 3)

Copyright

© 2021, Liu et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,884
    views
  • 394
    downloads
  • 18
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Lei Liu
  2. Zhiran Fan
  3. Xavier Rovira
  4. Li Xue
  5. Salomé Roux
  6. Isabelle Brabet
  7. Mingxia Xin
  8. Jean-Philippe Pin
  9. Jianfeng Liu
  10. Philippe Rondard
(2021)
Allosteric ligands control the activation of a class C GPCR heterodimer by acting at the transmembrane interface
eLife 10:e70188.
https://doi.org/10.7554/eLife.70188

Share this article

https://doi.org/10.7554/eLife.70188

Further reading

    1. Biochemistry and Chemical Biology
    2. Neuroscience
    Maximilian Nagel, Marco Niestroj ... Marc Spehr
    Research Article

    In most mammals, conspecific chemosensory communication relies on semiochemical release within complex bodily secretions and subsequent stimulus detection by the vomeronasal organ (VNO). Urine, a rich source of ethologically relevant chemosignals, conveys detailed information about sex, social hierarchy, health, and reproductive state, which becomes accessible to a conspecific via vomeronasal sampling. So far, however, numerous aspects of social chemosignaling along the vomeronasal pathway remain unclear. Moreover, since virtually all research on vomeronasal physiology is based on secretions derived from inbred laboratory mice, it remains uncertain whether such stimuli provide a true representation of potentially more relevant cues found in the wild. Here, we combine a robust low-noise VNO activity assay with comparative molecular profiling of sex- and strain-specific mouse urine samples from two inbred laboratory strains as well as from wild mice. With comprehensive molecular portraits of these secretions, VNO activity analysis now enables us to (i) assess whether and, if so, how much sex/strain-selective ‘raw’ chemical information in urine is accessible via vomeronasal sampling; (ii) identify which chemicals exhibit sufficient discriminatory power to signal an animal’s sex, strain, or both; (iii) determine the extent to which wild mouse secretions are unique; and (iv) analyze whether vomeronasal response profiles differ between strains. We report both sex- and, in particular, strain-selective VNO representations of chemical information. Within the urinary ‘secretome’, both volatile compounds and proteins exhibit sufficient discriminative power to provide sex- and strain-specific molecular fingerprints. While total protein amount is substantially enriched in male urine, females secrete a larger variety at overall comparatively low concentrations. Surprisingly, the molecular spectrum of wild mouse urine does not dramatically exceed that of inbred strains. Finally, vomeronasal response profiles differ between C57BL/6 and BALB/c animals, with particularly disparate representations of female semiochemicals.

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics
    Claudia D Consalvo, Adedeji M Aderounmu ... Brenda L Bass
    Research Article

    Invertebrates use the endoribonuclease Dicer to cleave viral dsRNA during antiviral defense, while vertebrates use RIG-I-like Receptors (RLRs), which bind viral dsRNA to trigger an interferon response. While some invertebrate Dicers act alone during antiviral defense, Caenorhabditis elegans Dicer acts in a complex with a dsRNA binding protein called RDE-4, and an RLR ortholog called DRH-1. We used biochemical and structural techniques to provide mechanistic insight into how these proteins function together. We found RDE-4 is important for ATP-independent and ATP-dependent cleavage reactions, while helicase domains of both DCR-1 and DRH-1 contribute to ATP-dependent cleavage. DRH-1 plays the dominant role in ATP hydrolysis, and like mammalian RLRs, has an N-terminal domain that functions in autoinhibition. A cryo-EM structure indicates DRH-1 interacts with DCR-1’s helicase domain, suggesting this interaction relieves autoinhibition. Our study unravels the mechanistic basis of the collaboration between two helicases from typically distinct innate immune defense pathways.