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Tracking receptor motions at the plasma membrane reveals distinct effects of ligands on CCR5 dynamics depending on its dimerization status
G-protein-coupled receptors (GPCR) are present at the cell surface in different conformational and oligomeric states. However, how these states impact GPCRs biological function and therapeutic targeting remains incompletely known. Here, we investigated this issue in living cells for the CC chemokine...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9307273/ https://www.ncbi.nlm.nih.gov/pubmed/35866628 http://dx.doi.org/10.7554/eLife.76281 |
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author | Momboisse, Fanny Nardi, Giacomo Colin, Philippe Hery, Melanie Cordeiro, Nelia Blachier, Simon Schwartz, Olivier Arenzana-Seisdedos, Fernando Sauvonnet, Nathalie Olivo-Marin, Jean-Christophe Lagane, Bernard Lagache, Thibault Brelot, Anne |
author_facet | Momboisse, Fanny Nardi, Giacomo Colin, Philippe Hery, Melanie Cordeiro, Nelia Blachier, Simon Schwartz, Olivier Arenzana-Seisdedos, Fernando Sauvonnet, Nathalie Olivo-Marin, Jean-Christophe Lagane, Bernard Lagache, Thibault Brelot, Anne |
author_sort | Momboisse, Fanny |
collection | PubMed |
description | G-protein-coupled receptors (GPCR) are present at the cell surface in different conformational and oligomeric states. However, how these states impact GPCRs biological function and therapeutic targeting remains incompletely known. Here, we investigated this issue in living cells for the CC chemokine receptor 5 (CCR5), a major receptor in inflammation and the principal entry co-receptor for Human Immunodeficiency Viruses type 1 (HIV-1). We used TIRF microscopy and a statistical method to track and classify the motion of different receptor subpopulations. We showed a diversity of ligand-free forms of CCR5 at the cell surface constituted of various oligomeric states and exhibiting transient Brownian and restricted motions. These forms were stabilized differently by distinct ligands. In particular, agonist stimulation restricted the mobility of CCR5 and led to its clustering, a feature depending on β-arrestin, while inverse agonist stimulation exhibited the opposite effect. These results suggest a link between receptor activation and immobilization. Applied to HIV-1 envelope glycoproteins gp120, our quantitative analysis revealed agonist-like properties of gp120s. Distinct gp120s influenced CCR5 dynamics differently, suggesting that they stabilize different CCR5 conformations. Then, using a dimerization-compromized mutant, we showed that dimerization (i) impacts CCR5 precoupling to G proteins, (ii) is a pre-requisite for the immobilization and clustering of receptors upon activation, and (iii) regulates receptor endocytosis, thereby impacting the fate of activated receptors. This study demonstrates that tracking the dynamic behavior of a GPCR is an efficient way to link GPCR conformations to their functions, therefore improving the development of drugs targeting specific receptor conformations. |
format | Online Article Text |
id | pubmed-9307273 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-93072732022-07-23 Tracking receptor motions at the plasma membrane reveals distinct effects of ligands on CCR5 dynamics depending on its dimerization status Momboisse, Fanny Nardi, Giacomo Colin, Philippe Hery, Melanie Cordeiro, Nelia Blachier, Simon Schwartz, Olivier Arenzana-Seisdedos, Fernando Sauvonnet, Nathalie Olivo-Marin, Jean-Christophe Lagane, Bernard Lagache, Thibault Brelot, Anne eLife Cell Biology G-protein-coupled receptors (GPCR) are present at the cell surface in different conformational and oligomeric states. However, how these states impact GPCRs biological function and therapeutic targeting remains incompletely known. Here, we investigated this issue in living cells for the CC chemokine receptor 5 (CCR5), a major receptor in inflammation and the principal entry co-receptor for Human Immunodeficiency Viruses type 1 (HIV-1). We used TIRF microscopy and a statistical method to track and classify the motion of different receptor subpopulations. We showed a diversity of ligand-free forms of CCR5 at the cell surface constituted of various oligomeric states and exhibiting transient Brownian and restricted motions. These forms were stabilized differently by distinct ligands. In particular, agonist stimulation restricted the mobility of CCR5 and led to its clustering, a feature depending on β-arrestin, while inverse agonist stimulation exhibited the opposite effect. These results suggest a link between receptor activation and immobilization. Applied to HIV-1 envelope glycoproteins gp120, our quantitative analysis revealed agonist-like properties of gp120s. Distinct gp120s influenced CCR5 dynamics differently, suggesting that they stabilize different CCR5 conformations. Then, using a dimerization-compromized mutant, we showed that dimerization (i) impacts CCR5 precoupling to G proteins, (ii) is a pre-requisite for the immobilization and clustering of receptors upon activation, and (iii) regulates receptor endocytosis, thereby impacting the fate of activated receptors. This study demonstrates that tracking the dynamic behavior of a GPCR is an efficient way to link GPCR conformations to their functions, therefore improving the development of drugs targeting specific receptor conformations. eLife Sciences Publications, Ltd 2022-07-22 /pmc/articles/PMC9307273/ /pubmed/35866628 http://dx.doi.org/10.7554/eLife.76281 Text en © 2022, Momboisse et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Cell Biology Momboisse, Fanny Nardi, Giacomo Colin, Philippe Hery, Melanie Cordeiro, Nelia Blachier, Simon Schwartz, Olivier Arenzana-Seisdedos, Fernando Sauvonnet, Nathalie Olivo-Marin, Jean-Christophe Lagane, Bernard Lagache, Thibault Brelot, Anne Tracking receptor motions at the plasma membrane reveals distinct effects of ligands on CCR5 dynamics depending on its dimerization status |
title | Tracking receptor motions at the plasma membrane reveals distinct effects of ligands on CCR5 dynamics depending on its dimerization status |
title_full | Tracking receptor motions at the plasma membrane reveals distinct effects of ligands on CCR5 dynamics depending on its dimerization status |
title_fullStr | Tracking receptor motions at the plasma membrane reveals distinct effects of ligands on CCR5 dynamics depending on its dimerization status |
title_full_unstemmed | Tracking receptor motions at the plasma membrane reveals distinct effects of ligands on CCR5 dynamics depending on its dimerization status |
title_short | Tracking receptor motions at the plasma membrane reveals distinct effects of ligands on CCR5 dynamics depending on its dimerization status |
title_sort | tracking receptor motions at the plasma membrane reveals distinct effects of ligands on ccr5 dynamics depending on its dimerization status |
topic | Cell Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9307273/ https://www.ncbi.nlm.nih.gov/pubmed/35866628 http://dx.doi.org/10.7554/eLife.76281 |
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