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Generation of Gαi knock-out HEK293 cells illuminates Gαi-coupling diversity of GPCRs

G-protein-coupled receptors (GPCRs) are pivotal cell membrane proteins that sense extracellular molecules and activate cellular responses. The G-protein α subunit i (Gαi) family represents the most common GPCR-coupling partner and consists of eight subunits with distinct signaling properties. Howeve...

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Detalles Bibliográficos
Autores principales: Ono, Yuki, Kawakami, Kouki, Nakamura, Gaku, Ishida, Satoru, Aoki, Junken, Inoue, Asuka
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9884212/
https://www.ncbi.nlm.nih.gov/pubmed/36709222
http://dx.doi.org/10.1038/s42003-023-04465-2
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author Ono, Yuki
Kawakami, Kouki
Nakamura, Gaku
Ishida, Satoru
Aoki, Junken
Inoue, Asuka
author_facet Ono, Yuki
Kawakami, Kouki
Nakamura, Gaku
Ishida, Satoru
Aoki, Junken
Inoue, Asuka
author_sort Ono, Yuki
collection PubMed
description G-protein-coupled receptors (GPCRs) are pivotal cell membrane proteins that sense extracellular molecules and activate cellular responses. The G-protein α subunit i (Gαi) family represents the most common GPCR-coupling partner and consists of eight subunits with distinct signaling properties. However, analyzing the coupling pattern has been challenging owing to endogenous expression of the Gαi subunits in virtually all cell lines. Here, we generate a HEK293 cell line lacking all Gαi subunits, which enables the measurement of GPCR-Gαi coupling upon transient re-expression of a specific Gαi subunit. We profile Gαi-coupling selectivity across 11 GPCRs by measuring ligand-induced inhibitory activity for cAMP accumulation. The coupling profiles are then classified into three clusters, representing those preferentially coupled to Gαz, those to Gαo, and those with unapparent selectivity. These results indicate that individual Gαi-coupled GPCRs fine-tune Gαi signaling by exerting coupling preference at the Gαi-subunit level.
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spelling pubmed-98842122023-01-30 Generation of Gαi knock-out HEK293 cells illuminates Gαi-coupling diversity of GPCRs Ono, Yuki Kawakami, Kouki Nakamura, Gaku Ishida, Satoru Aoki, Junken Inoue, Asuka Commun Biol Article G-protein-coupled receptors (GPCRs) are pivotal cell membrane proteins that sense extracellular molecules and activate cellular responses. The G-protein α subunit i (Gαi) family represents the most common GPCR-coupling partner and consists of eight subunits with distinct signaling properties. However, analyzing the coupling pattern has been challenging owing to endogenous expression of the Gαi subunits in virtually all cell lines. Here, we generate a HEK293 cell line lacking all Gαi subunits, which enables the measurement of GPCR-Gαi coupling upon transient re-expression of a specific Gαi subunit. We profile Gαi-coupling selectivity across 11 GPCRs by measuring ligand-induced inhibitory activity for cAMP accumulation. The coupling profiles are then classified into three clusters, representing those preferentially coupled to Gαz, those to Gαo, and those with unapparent selectivity. These results indicate that individual Gαi-coupled GPCRs fine-tune Gαi signaling by exerting coupling preference at the Gαi-subunit level. Nature Publishing Group UK 2023-01-28 /pmc/articles/PMC9884212/ /pubmed/36709222 http://dx.doi.org/10.1038/s42003-023-04465-2 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Ono, Yuki
Kawakami, Kouki
Nakamura, Gaku
Ishida, Satoru
Aoki, Junken
Inoue, Asuka
Generation of Gαi knock-out HEK293 cells illuminates Gαi-coupling diversity of GPCRs
title Generation of Gαi knock-out HEK293 cells illuminates Gαi-coupling diversity of GPCRs
title_full Generation of Gαi knock-out HEK293 cells illuminates Gαi-coupling diversity of GPCRs
title_fullStr Generation of Gαi knock-out HEK293 cells illuminates Gαi-coupling diversity of GPCRs
title_full_unstemmed Generation of Gαi knock-out HEK293 cells illuminates Gαi-coupling diversity of GPCRs
title_short Generation of Gαi knock-out HEK293 cells illuminates Gαi-coupling diversity of GPCRs
title_sort generation of gαi knock-out hek293 cells illuminates gαi-coupling diversity of gpcrs
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9884212/
https://www.ncbi.nlm.nih.gov/pubmed/36709222
http://dx.doi.org/10.1038/s42003-023-04465-2
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