Effector membrane translocation biosensors reveal G protein and βarrestin coupling profiles of 100 therapeutically relevant GPCRs
- University of Montreal, Canada
- Domain Therapeutics North America, Canada
- University of Copenhagen, Denmark
- Pfizer Worldwide Research, United States
- Pfizer Global R&D, United States
- 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.
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G protein selectivity - a unified meta-analysisAccompanying 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
Alexander Sebastian Hauser
Hiroyuki Kobayashi
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.
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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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
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