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

Charlotte Avet; Arturo Mancini; Billy Breton; Christian Le Gouill; Alexander S. Hauser; Claire Normand; Hiroyuki Kobayashi; Florence Gross; Mireille Hogue; Viktoriya Lukasheva; Stéphane St-Onge; Marilyn Carrier; Madeleine Héroux; Sandra Morissette; Eric B. Fauman; Jean Philippe Fortin; Stephan Schann; Xavier Leroy; David E. Gloriam; Michel Bouvier

doi:10.7554/eLife.74101

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

Charlotte Avet, Arturo Mancini, Billy Breton, Christian Le Gouill, Alexander S. Hauser, Claire Normand, Hiroyuki Kobayashi, Florence Gross, Mireille Hogue, Viktoriya Lukasheva, Stéphane St-Onge, Marilyn Carrier, Madeleine Héroux, Sandra Morissette, Eric B. Fauman, Jean Philippe Fortin, Stephan Schann, Xavier Leroy, David E. Gloriam, Michel Bouvier

Biopharmaceuticals

Research output: Contribution to journal › Journal article › Research › peer-review

130 Citations (Scopus)

19 Downloads (Pure)

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 βarrestins 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 1500 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.

Original language	English
Article number	e74101
Journal	eLife
Volume	11
Number of pages	34
ISSN	2050-084X
DOIs	https://doi.org/10.7554/eLife.74101
Publication status	Published - 2022

Bibliographical note

Publisher Copyright:
© 2022, Avet et al.

Keywords

biochemistry
biosensor
chemical biology
effector membrane translocation assay
enhanced bystander bioluminescence resonance energy transfer
G protein activation
g protein-coupled receptor
high-throughput assay
human

Access to Document

10.7554/eLife.74101

FulltextFinal published version, 4.86 MBLicence: CC BY

Cite this

Avet, C., Mancini, A., Breton, B., Le Gouill, C., Hauser, A. S., Normand, C., Kobayashi, H., Gross, F., Hogue, M., Lukasheva, V., St-Onge, S., Carrier, M., Héroux, M., Morissette, S., Fauman, E. B., Fortin, J. P., Schann, S., Leroy, X., Gloriam, D. E., & Bouvier, M. (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

Effector membrane translocation biosensors reveal G protein and βarrestin coupling profiles of 100 therapeutically relevant GPCRs. / Avet, Charlotte; Mancini, Arturo; Breton, Billy; Le Gouill, Christian; Hauser, Alexander S.; Normand, Claire; Kobayashi, Hiroyuki; Gross, Florence; Hogue, Mireille; Lukasheva, Viktoriya; St-Onge, Stéphane; Carrier, Marilyn; Héroux, Madeleine; Morissette, Sandra; Fauman, Eric B.; Fortin, Jean Philippe; Schann, Stephan; Leroy, Xavier; Gloriam, David E.; Bouvier, Michel.

In: eLife, Vol. 11, e74101, 2022.

Research output: Contribution to journal › Journal article › Research › peer-review

Avet, C, Mancini, A, Breton, B, Le Gouill, C, Hauser, AS, Normand, C, Kobayashi, H, Gross, F, Hogue, M, Lukasheva, V, St-Onge, S, Carrier, M, Héroux, M, Morissette, S, Fauman, EB, Fortin, JP, Schann, S, Leroy, X, Gloriam, DE & Bouvier, M 2022, 'Effector membrane translocation biosensors reveal G protein and βarrestin coupling profiles of 100 therapeutically relevant GPCRs', eLife, vol. 11, e74101. https://doi.org/10.7554/eLife.74101

Avet, Charlotte ; Mancini, Arturo ; Breton, Billy ; Le Gouill, Christian ; Hauser, Alexander S. ; Normand, Claire ; Kobayashi, Hiroyuki ; Gross, Florence ; Hogue, Mireille ; Lukasheva, Viktoriya ; St-Onge, Stéphane ; Carrier, Marilyn ; Héroux, Madeleine ; Morissette, Sandra ; Fauman, Eric B. ; Fortin, Jean Philippe ; Schann, Stephan ; Leroy, Xavier ; Gloriam, David E. ; Bouvier, Michel. / Effector membrane translocation biosensors reveal G protein and βarrestin coupling profiles of 100 therapeutically relevant GPCRs. In: eLife. 2022 ; Vol. 11.

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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 βarrestins 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 1500 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.",

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