Cargando…
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...
Autores principales: | , , , |
---|---|
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 |
_version_ | 1783681680270163968 |
---|---|
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. |
format | Online Article Text |
id | pubmed-8062009 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT lusumin constitutivegproteincouplingprofilesofunderstudiedorphangpcrs AT jangwonjo constitutivegproteincouplingprofilesofunderstudiedorphangpcrs AT inoueasuka constitutivegproteincouplingprofilesofunderstudiedorphangpcrs AT lambertnevina constitutivegproteincouplingprofilesofunderstudiedorphangpcrs |