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Mass spectrometry captures biased signalling and allosteric modulation of a G-protein-coupled receptor
G-protein-coupled receptors signal through cognate G proteins. Despite the widespread importance of these receptors, their regulatory mechanisms for G-protein selectivity are not fully understood. Here we present a native mass spectrometry-based approach to interrogate both biased signalling and all...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9758051/ https://www.ncbi.nlm.nih.gov/pubmed/36357787 http://dx.doi.org/10.1038/s41557-022-01041-9 |
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author | Yen, Hsin-Yung Liko, Idlir Song, Wanling Kapoor, Parth Almeida, Fernando Toporowska, Joanna Gherbi, Karolina Hopper, Jonathan T. S. Charlton, Steven J. Politis, Argyris Sansom, Mark S. P. Jazayeri, Ali Robinson, Carol V. |
author_facet | Yen, Hsin-Yung Liko, Idlir Song, Wanling Kapoor, Parth Almeida, Fernando Toporowska, Joanna Gherbi, Karolina Hopper, Jonathan T. S. Charlton, Steven J. Politis, Argyris Sansom, Mark S. P. Jazayeri, Ali Robinson, Carol V. |
author_sort | Yen, Hsin-Yung |
collection | PubMed |
description | G-protein-coupled receptors signal through cognate G proteins. Despite the widespread importance of these receptors, their regulatory mechanisms for G-protein selectivity are not fully understood. Here we present a native mass spectrometry-based approach to interrogate both biased signalling and allosteric modulation of the β(1)-adrenergic receptor in response to various ligands. By simultaneously capturing the effects of ligand binding and receptor coupling to different G proteins, we probed the relative importance of specific interactions with the receptor through systematic changes in 14 ligands, including isoprenaline derivatives, full and partial agonists, and antagonists. We observed enhanced dynamics of the intracellular loop 3 in the presence of isoprenaline, which is capable of acting as a biased agonist. We also show here that endogenous zinc ions augment the binding in receptor–G(s) complexes and propose a zinc ion-binding hotspot at the TM5/TM6 intracellular interface of the receptor–G(s) complex. Further interrogation led us to propose a mechanism in which zinc ions facilitate a structural transition of the intermediate complex towards the stable state. [Image: see text] |
format | Online Article Text |
id | pubmed-9758051 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-97580512022-12-18 Mass spectrometry captures biased signalling and allosteric modulation of a G-protein-coupled receptor Yen, Hsin-Yung Liko, Idlir Song, Wanling Kapoor, Parth Almeida, Fernando Toporowska, Joanna Gherbi, Karolina Hopper, Jonathan T. S. Charlton, Steven J. Politis, Argyris Sansom, Mark S. P. Jazayeri, Ali Robinson, Carol V. Nat Chem Article G-protein-coupled receptors signal through cognate G proteins. Despite the widespread importance of these receptors, their regulatory mechanisms for G-protein selectivity are not fully understood. Here we present a native mass spectrometry-based approach to interrogate both biased signalling and allosteric modulation of the β(1)-adrenergic receptor in response to various ligands. By simultaneously capturing the effects of ligand binding and receptor coupling to different G proteins, we probed the relative importance of specific interactions with the receptor through systematic changes in 14 ligands, including isoprenaline derivatives, full and partial agonists, and antagonists. We observed enhanced dynamics of the intracellular loop 3 in the presence of isoprenaline, which is capable of acting as a biased agonist. We also show here that endogenous zinc ions augment the binding in receptor–G(s) complexes and propose a zinc ion-binding hotspot at the TM5/TM6 intracellular interface of the receptor–G(s) complex. Further interrogation led us to propose a mechanism in which zinc ions facilitate a structural transition of the intermediate complex towards the stable state. [Image: see text] Nature Publishing Group UK 2022-11-10 2022 /pmc/articles/PMC9758051/ /pubmed/36357787 http://dx.doi.org/10.1038/s41557-022-01041-9 Text en © The Author(s) 2022 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 Yen, Hsin-Yung Liko, Idlir Song, Wanling Kapoor, Parth Almeida, Fernando Toporowska, Joanna Gherbi, Karolina Hopper, Jonathan T. S. Charlton, Steven J. Politis, Argyris Sansom, Mark S. P. Jazayeri, Ali Robinson, Carol V. Mass spectrometry captures biased signalling and allosteric modulation of a G-protein-coupled receptor |
title | Mass spectrometry captures biased signalling and allosteric modulation of a G-protein-coupled receptor |
title_full | Mass spectrometry captures biased signalling and allosteric modulation of a G-protein-coupled receptor |
title_fullStr | Mass spectrometry captures biased signalling and allosteric modulation of a G-protein-coupled receptor |
title_full_unstemmed | Mass spectrometry captures biased signalling and allosteric modulation of a G-protein-coupled receptor |
title_short | Mass spectrometry captures biased signalling and allosteric modulation of a G-protein-coupled receptor |
title_sort | mass spectrometry captures biased signalling and allosteric modulation of a g-protein-coupled receptor |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9758051/ https://www.ncbi.nlm.nih.gov/pubmed/36357787 http://dx.doi.org/10.1038/s41557-022-01041-9 |
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