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Conformational plasticity of ligand-bound and ternary GPCR complexes studied by (19)F NMR of the β(1)-adrenergic receptor
G-protein-coupled receptors (GPCRs) are allosteric signaling proteins that transmit an extracellular stimulus across the cell membrane. Using (19)F NMR and site-specific labelling, we investigate the response of the cytoplasmic region of transmembrane helices 6 and 7 of the β(1)-adrenergic receptor...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6997182/ https://www.ncbi.nlm.nih.gov/pubmed/32015348 http://dx.doi.org/10.1038/s41467-020-14526-3 |
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author | Frei, J. Niclas Broadhurst, Richard W. Bostock, Mark J. Solt, Andras Jones, Andrew J. Y. Gabriel, Florian Tandale, Aditi Shrestha, Binesh Nietlispach, Daniel |
author_facet | Frei, J. Niclas Broadhurst, Richard W. Bostock, Mark J. Solt, Andras Jones, Andrew J. Y. Gabriel, Florian Tandale, Aditi Shrestha, Binesh Nietlispach, Daniel |
author_sort | Frei, J. Niclas |
collection | PubMed |
description | G-protein-coupled receptors (GPCRs) are allosteric signaling proteins that transmit an extracellular stimulus across the cell membrane. Using (19)F NMR and site-specific labelling, we investigate the response of the cytoplasmic region of transmembrane helices 6 and 7 of the β(1)-adrenergic receptor to agonist stimulation and coupling to a G(s)-protein-mimetic nanobody. Agonist binding shows the receptor in equilibrium between two inactive states and a pre-active form, increasingly populated with higher ligand efficacy. Nanobody coupling leads to a fully active ternary receptor complex present in amounts correlating directly with agonist efficacy, consistent with partial agonism. While for different agonists the helix 6 environment in the active-state ternary complexes resides in a well-defined conformation, showing little conformational mobility, the environment of the highly conserved NPxxY motif on helix 7 remains dynamic adopting diverse, agonist-specific conformations, implying a further role of this region in receptor function. An inactive nanobody-coupled ternary receptor form is also observed. |
format | Online Article Text |
id | pubmed-6997182 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-69971822020-02-05 Conformational plasticity of ligand-bound and ternary GPCR complexes studied by (19)F NMR of the β(1)-adrenergic receptor Frei, J. Niclas Broadhurst, Richard W. Bostock, Mark J. Solt, Andras Jones, Andrew J. Y. Gabriel, Florian Tandale, Aditi Shrestha, Binesh Nietlispach, Daniel Nat Commun Article G-protein-coupled receptors (GPCRs) are allosteric signaling proteins that transmit an extracellular stimulus across the cell membrane. Using (19)F NMR and site-specific labelling, we investigate the response of the cytoplasmic region of transmembrane helices 6 and 7 of the β(1)-adrenergic receptor to agonist stimulation and coupling to a G(s)-protein-mimetic nanobody. Agonist binding shows the receptor in equilibrium between two inactive states and a pre-active form, increasingly populated with higher ligand efficacy. Nanobody coupling leads to a fully active ternary receptor complex present in amounts correlating directly with agonist efficacy, consistent with partial agonism. While for different agonists the helix 6 environment in the active-state ternary complexes resides in a well-defined conformation, showing little conformational mobility, the environment of the highly conserved NPxxY motif on helix 7 remains dynamic adopting diverse, agonist-specific conformations, implying a further role of this region in receptor function. An inactive nanobody-coupled ternary receptor form is also observed. Nature Publishing Group UK 2020-02-03 /pmc/articles/PMC6997182/ /pubmed/32015348 http://dx.doi.org/10.1038/s41467-020-14526-3 Text en © The Author(s) 2020 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/. |
spellingShingle | Article Frei, J. Niclas Broadhurst, Richard W. Bostock, Mark J. Solt, Andras Jones, Andrew J. Y. Gabriel, Florian Tandale, Aditi Shrestha, Binesh Nietlispach, Daniel Conformational plasticity of ligand-bound and ternary GPCR complexes studied by (19)F NMR of the β(1)-adrenergic receptor |
title | Conformational plasticity of ligand-bound and ternary GPCR complexes studied by (19)F NMR of the β(1)-adrenergic receptor |
title_full | Conformational plasticity of ligand-bound and ternary GPCR complexes studied by (19)F NMR of the β(1)-adrenergic receptor |
title_fullStr | Conformational plasticity of ligand-bound and ternary GPCR complexes studied by (19)F NMR of the β(1)-adrenergic receptor |
title_full_unstemmed | Conformational plasticity of ligand-bound and ternary GPCR complexes studied by (19)F NMR of the β(1)-adrenergic receptor |
title_short | Conformational plasticity of ligand-bound and ternary GPCR complexes studied by (19)F NMR of the β(1)-adrenergic receptor |
title_sort | conformational plasticity of ligand-bound and ternary gpcr complexes studied by (19)f nmr of the β(1)-adrenergic receptor |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6997182/ https://www.ncbi.nlm.nih.gov/pubmed/32015348 http://dx.doi.org/10.1038/s41467-020-14526-3 |
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