Effector membrane translocation biosensors reveal G protein and βarrestin coupling profiles of 100 therapeutically relevant GPCRs

  1. Charlotte Avet
  2. Arturo Mancini
  3. Billy Breton
  4. Christian Le Gouill
  5. Alexander Sebastian Hauser
  6. Claire Normand
  7. Hiroyuki Kobayashi
  8. Florence Gross
  9. Mireille Hogue
  10. Viktoriya Lukasheva
  11. Stéphane St-Onge
  12. Marilyn Carrier
  13. Madeleine Héroux
  14. Sandra Morissette
  15. Eric B Fauman
  16. Jean-Philippe Fortin
  17. Stephan Schann
  18. Xavier Leroy  Is a corresponding author
  19. David E Gloriam  Is a corresponding author
  20. Michel Bouvier  Is a corresponding author
  1. University of Montreal, Canada
  2. Domain Therapeutics North America, Canada
  3. University of Copenhagen, Denmark
  4. Pfizer Worldwide Research, United States
  5. Pfizer Global R&D, United States
  6. Domain Therapeutics, France

Abstract

The recognition that individual GPCRs can activate multiple signaling pathways has raised the possibility of developing drugs selectively targeting therapeutically relevant ones. This requires tools to determine which G proteins and barrestins are activated by a given receptor. Here, we present a set of BRET sensors monitoring the activation of the 12 G protein subtypes based on the translocation of their effectors to the plasma membrane (EMTA). Unlike most of the existing detection systems, EMTA does not require modification of receptors or G proteins (except for Gs). EMTA was found to be suitable for the detection of constitutive activity, inverse agonism, biased signaling and polypharmacology. Profiling of 100 therapeutically relevant human GPCRs resulted in 1,500 pathway-specific concentration-response curves and revealed a great diversity of coupling profiles ranging from exquisite selectivity to broad promiscuity. Overall, this work describes unique resources for studying the complexities underlying GPCR signaling and pharmacology.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for Figures 2, 4, 5, 6 and 7 and associated figure supplements. Supplementary Table 1 contains the numerical data used to generate Figure 2-figure supplement 1 and Figure 2-figure supplement 3. Supplementary Table 2 contains the numerical data used to generate figure 3.

The following data sets were generated
    1. Hauser AS
    2. Avet C
    3. Normand C
    4. Mancini A
    5. Inoue A
    6. Bouvier M
    7. Gloriam DE
    (2021) G protein selectivity - a unified meta-analysis
    Accompanying dataset paper in BioRXiv (https://doi.org/10.1101/2021.09.07.459250) and www.gpcrdb.org (https://doi.org/10.1093/nar/gkab852).

Article and author information

Author details

  1. Charlotte Avet

    Department of Biochemistry and Molecular Medicine, University of Montreal, Montréal, Canada
    Competing interests
    No competing interests declared.
  2. Arturo Mancini

    Domain Therapeutics North America, Montréal, Canada
    Competing interests
    Arturo Mancini, was employee of Domain Therapeutics North America during this research.
  3. Billy Breton

    Domain Therapeutics North America, Montréal, Canada
    Competing interests
    Billy Breton, was employee of Domain Therapeutics North America during this research. Has filed patent application (US20200256869A1) related to the biosensors used in this work and the technology has been licensed to Domain Therapeutics..
  4. Christian Le Gouill

    Department of Biochemistry and Molecular Medicine, University of Montreal, Montréal, Canada
    Competing interests
    Christian Le Gouill, has filed patent application (US20200256869A1) related to the biosensors used in this work and the technology has been licensed to Domain Therapeutics..
  5. Alexander Sebastian Hauser

    Department of Drug Design and Pharmacology, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1098-6419
  6. Claire Normand

    Domain Therapeutics North America, Montréal, Canada
    Competing interests
    Claire Normand, was employee of Domain Therapeutics North America during this research.
  7. Hiroyuki Kobayashi

    Department of Biochemistry and Molecular Medicine, University of Montreal, Montréal, Canada
    Competing interests
    Hiroyuki Kobayashi, has filed patent application (US20200256869A1) related to the biosensors used in this work and the technology has been licensed to Domain Therapeutics..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4965-0883
  8. Florence Gross

    Domain Therapeutics North America, Montréal, Canada
    Competing interests
    Florence Gross, was employee of Domain Therapeutics North America during this research.
  9. Mireille Hogue

    Department of Biochemistry and Molecular Medicine, University of Montreal, Montréal, Canada
    Competing interests
    Mireille Hogue, has filed patent application (US20200256869A1) related to the biosensors used in this work and the technology has been licensed to Domain Therapeutics..
  10. Viktoriya Lukasheva

    Department of Biochemistry and Molecular Medicine, University of Montreal, Montréal, Canada
    Competing interests
    Viktoriya Lukasheva, has filed patent application (US20200256869A1) related to the biosensors used in this work and the technology has been licensed to Domain Therapeutics..
  11. Stéphane St-Onge

    Department of Biochemistry and Molecular Medicine, University of Montreal, Montréal, Canada
    Competing interests
    No competing interests declared.
  12. Marilyn Carrier

    Department of Biochemistry and Molecular Medicine, University of Montreal, Montréal, Canada
    Competing interests
    No competing interests declared.
  13. Madeleine Héroux

    Department of Biochemistry and Molecular Medicine, University of Montreal, Montréal, Canada
    Competing interests
    No competing interests declared.
  14. Sandra Morissette

    Domain Therapeutics North America, Montréal, Canada
    Competing interests
    Sandra Morissette, SM was employee of Domain Therapeutics North America during this research.
  15. Eric B Fauman

    Internal Medicine Research Unit, Pfizer Worldwide Research, Cambridge, United States
    Competing interests
    Eric B Fauman, is employee and shares holders of Pfizer.
  16. Jean-Philippe Fortin

    Pfizer Global R&D, Cambridge, United States
    Competing interests
    Jean-Philippe Fortin, is employee and shares holders of Pfizer.
  17. Stephan Schann

    Domain Therapeutics, Illkirch-Strasbourg, France
    Competing interests
    Stephan Schann, is employee and is part of the management of Domain Therapeutics.
  18. Xavier Leroy

    Domain Therapeutics, llkirch-Strasbourg, France
    For correspondence
    xleroy@domaintherapeutics.com
    Competing interests
    Xavier Leroy, is employee and is part of the management of Domain Therapeutics.
  19. David E Gloriam

    Department of Drug Design and Pharmacology, University of Copenhagen, Copenhagen, Denmark
    For correspondence
    david.gloriam@sund.ku.dk
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4299-7561
  20. Michel Bouvier

    Department of Biochemistry and Molecular Medicine, University of Montreal, Montréal, Canada
    For correspondence
    michel.bouvier@umontreal.ca
    Competing interests
    Michel Bouvier, is the president of Domain Therapeutics scientific Has filed patent application (US20200256869A1) related to the biosensors used in this work and the technology has been licensed to Domain Therapeutics..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1128-0100

Funding

Canada Research Chairs

  • Michel Bouvier

Canadian Institutes of Health Research (FDN-148431)

  • Michel Bouvier

Lundbeckfonden (R218-2016-1266)

  • David E Gloriam

Lundbeckfonden (R313-2019-526)

  • David E Gloriam

Novo Nordisk Fonden (NNF18OC0031226)

  • David E Gloriam

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

Reviewing Editor

  1. William I Weis, Stanford University School of Medicine, United States

Version history

  1. Preprint posted: April 24, 2020 (view preprint)
  2. Received: September 21, 2021
  3. Accepted: March 17, 2022
  4. Accepted Manuscript published: March 18, 2022 (version 1)
  5. Version of Record published: April 12, 2022 (version 2)

Copyright

© 2022, Avet 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.

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  1. Charlotte Avet
  2. Arturo Mancini
  3. Billy Breton
  4. Christian Le Gouill
  5. Alexander Sebastian Hauser
  6. Claire Normand
  7. Hiroyuki Kobayashi
  8. Florence Gross
  9. Mireille Hogue
  10. Viktoriya Lukasheva
  11. Stéphane St-Onge
  12. Marilyn Carrier
  13. Madeleine Héroux
  14. Sandra Morissette
  15. Eric B Fauman
  16. Jean-Philippe Fortin
  17. Stephan Schann
  18. Xavier Leroy
  19. David E Gloriam
  20. Michel Bouvier
(2022)
Effector membrane translocation biosensors reveal G protein and βarrestin coupling profiles of 100 therapeutically relevant GPCRs
eLife 11:e74101.
https://doi.org/10.7554/eLife.74101

Share this article

https://doi.org/10.7554/eLife.74101

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