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Constitutive G protein coupling profiles of understudied orphan GPCRs

A large number of GPCRs are potentially valuable drug targets but remain understudied. Many of these lack well-validated activating ligands and are considered “orphan” receptors, and G protein coupling profiles have not been defined for many orphan GPCRs. Here we asked if constitutive receptor activ...

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Detalles Bibliográficos
Autores principales: Lu, Sumin, Jang, Wonjo, Inoue, Asuka, Lambert, Nevin A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8062009/
https://www.ncbi.nlm.nih.gov/pubmed/33886554
http://dx.doi.org/10.1371/journal.pone.0247743
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author Lu, Sumin
Jang, Wonjo
Inoue, Asuka
Lambert, Nevin A.
author_facet Lu, Sumin
Jang, Wonjo
Inoue, Asuka
Lambert, Nevin A.
author_sort Lu, Sumin
collection PubMed
description A large number of GPCRs are potentially valuable drug targets but remain understudied. Many of these lack well-validated activating ligands and are considered “orphan” receptors, and G protein coupling profiles have not been defined for many orphan GPCRs. Here we asked if constitutive receptor activity can be used to determine G protein coupling profiles of orphan GPCRs. We monitored nucleotide-sensitive interactions between 48 understudied orphan GPCRs and five G proteins (240 combinations) using bioluminescence resonance energy transfer (BRET). No receptor ligands were used, but GDP was used as a common G protein ligand to disrupt receptor-G protein complexes. Constitutive BRET between the same receptors and β-arrestins was also measured. We found sufficient GDP-sensitive BRET to generate G protein coupling profiles for 22 of the 48 receptors we studied. Altogether we identified 48 coupled receptor-G protein pairs, many of which have not been described previously. We conclude that receptor-G protein complexes that form spontaneously in the absence of guanine nucleotides can be used to profile G protein coupling of constitutively-active GPCRs. This approach may prove useful for studying G protein coupling of other GPCRs for which activating ligands are not available.
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spelling pubmed-80620092021-05-04 Constitutive G protein coupling profiles of understudied orphan GPCRs Lu, Sumin Jang, Wonjo Inoue, Asuka Lambert, Nevin A. PLoS One Research Article A large number of GPCRs are potentially valuable drug targets but remain understudied. Many of these lack well-validated activating ligands and are considered “orphan” receptors, and G protein coupling profiles have not been defined for many orphan GPCRs. Here we asked if constitutive receptor activity can be used to determine G protein coupling profiles of orphan GPCRs. We monitored nucleotide-sensitive interactions between 48 understudied orphan GPCRs and five G proteins (240 combinations) using bioluminescence resonance energy transfer (BRET). No receptor ligands were used, but GDP was used as a common G protein ligand to disrupt receptor-G protein complexes. Constitutive BRET between the same receptors and β-arrestins was also measured. We found sufficient GDP-sensitive BRET to generate G protein coupling profiles for 22 of the 48 receptors we studied. Altogether we identified 48 coupled receptor-G protein pairs, many of which have not been described previously. We conclude that receptor-G protein complexes that form spontaneously in the absence of guanine nucleotides can be used to profile G protein coupling of constitutively-active GPCRs. This approach may prove useful for studying G protein coupling of other GPCRs for which activating ligands are not available. Public Library of Science 2021-04-22 /pmc/articles/PMC8062009/ /pubmed/33886554 http://dx.doi.org/10.1371/journal.pone.0247743 Text en © 2021 Lu et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Lu, Sumin
Jang, Wonjo
Inoue, Asuka
Lambert, Nevin A.
Constitutive G protein coupling profiles of understudied orphan GPCRs
title Constitutive G protein coupling profiles of understudied orphan GPCRs
title_full Constitutive G protein coupling profiles of understudied orphan GPCRs
title_fullStr Constitutive G protein coupling profiles of understudied orphan GPCRs
title_full_unstemmed Constitutive G protein coupling profiles of understudied orphan GPCRs
title_short Constitutive G protein coupling profiles of understudied orphan GPCRs
title_sort constitutive g protein coupling profiles of understudied orphan gpcrs
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8062009/
https://www.ncbi.nlm.nih.gov/pubmed/33886554
http://dx.doi.org/10.1371/journal.pone.0247743
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