Unravelling the mechanism of neurotensin recognition by neurotensin receptor 1
Abstract The conformational ensembles of G protein-coupled receptors (GPCRs) include inactive and active states. Spectroscopy techniques, including NMR, show that agonists, antagonists and other ligands shift the ensemble toward specific states depending on the pharmacological efficacy of the ligand...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-12-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-44010-7 |
| _version_ | 1797397885697916928 |
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| author | Kazem Asadollahi Sunnia Rajput Lazarus Andrew de Zhang Ching-Seng Ang Shuai Nie Nicholas A. Williamson Michael D. W. Griffin Ross A. D. Bathgate Daniel J. Scott Thomas R. Weikl Guy N. L. Jameson Paul R. Gooley |
| author_facet | Kazem Asadollahi Sunnia Rajput Lazarus Andrew de Zhang Ching-Seng Ang Shuai Nie Nicholas A. Williamson Michael D. W. Griffin Ross A. D. Bathgate Daniel J. Scott Thomas R. Weikl Guy N. L. Jameson Paul R. Gooley |
| author_sort | Kazem Asadollahi |
| collection | DOAJ |
| description | Abstract The conformational ensembles of G protein-coupled receptors (GPCRs) include inactive and active states. Spectroscopy techniques, including NMR, show that agonists, antagonists and other ligands shift the ensemble toward specific states depending on the pharmacological efficacy of the ligand. How receptors recognize ligands and the kinetic mechanism underlying this population shift is poorly understood. Here, we investigate the kinetic mechanism of neurotensin recognition by neurotensin receptor 1 (NTS1) using 19F-NMR, hydrogen-deuterium exchange mass spectrometry and stopped-flow fluorescence spectroscopy. Our results indicate slow-exchanging conformational heterogeneity on the extracellular surface of ligand-bound NTS1. Numerical analysis of the kinetic data of neurotensin binding to NTS1 shows that ligand recognition follows an induced-fit mechanism, in which conformational changes occur after neurotensin binding. This approach is applicable to other GPCRs to provide insight into the kinetic regulation of ligand recognition by GPCRs. |
| first_indexed | 2024-03-09T01:16:40Z |
| format | Article |
| id | doaj.art-8a32b17098464f42af0a3f2a34e66fbb |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-09T01:16:40Z |
| publishDate | 2023-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-8a32b17098464f42af0a3f2a34e66fbb2023-12-10T12:25:00ZengNature PortfolioNature Communications2041-17232023-12-0114111310.1038/s41467-023-44010-7Unravelling the mechanism of neurotensin recognition by neurotensin receptor 1Kazem Asadollahi0Sunnia Rajput1Lazarus Andrew de Zhang2Ching-Seng Ang3Shuai Nie4Nicholas A. Williamson5Michael D. W. Griffin6Ross A. D. Bathgate7Daniel J. Scott8Thomas R. Weikl9Guy N. L. Jameson10Paul R. Gooley11Department of Biochemistry and Pharmacology, University of MelbourneBio21 Molecular Science and Biotechnology Institute, University of MelbourneThe Florey, University of MelbourneBio21 Molecular Science and Biotechnology Institute, University of MelbourneBio21 Molecular Science and Biotechnology Institute, University of MelbourneBio21 Molecular Science and Biotechnology Institute, University of MelbourneDepartment of Biochemistry and Pharmacology, University of MelbourneDepartment of Biochemistry and Pharmacology, University of MelbourneDepartment of Biochemistry and Pharmacology, University of MelbourneDepartment of Biomolecular Systems, Max Planck Institute of Colloids and InterfacesBio21 Molecular Science and Biotechnology Institute, University of MelbourneDepartment of Biochemistry and Pharmacology, University of MelbourneAbstract The conformational ensembles of G protein-coupled receptors (GPCRs) include inactive and active states. Spectroscopy techniques, including NMR, show that agonists, antagonists and other ligands shift the ensemble toward specific states depending on the pharmacological efficacy of the ligand. How receptors recognize ligands and the kinetic mechanism underlying this population shift is poorly understood. Here, we investigate the kinetic mechanism of neurotensin recognition by neurotensin receptor 1 (NTS1) using 19F-NMR, hydrogen-deuterium exchange mass spectrometry and stopped-flow fluorescence spectroscopy. Our results indicate slow-exchanging conformational heterogeneity on the extracellular surface of ligand-bound NTS1. Numerical analysis of the kinetic data of neurotensin binding to NTS1 shows that ligand recognition follows an induced-fit mechanism, in which conformational changes occur after neurotensin binding. This approach is applicable to other GPCRs to provide insight into the kinetic regulation of ligand recognition by GPCRs.https://doi.org/10.1038/s41467-023-44010-7 |
| spellingShingle | Kazem Asadollahi Sunnia Rajput Lazarus Andrew de Zhang Ching-Seng Ang Shuai Nie Nicholas A. Williamson Michael D. W. Griffin Ross A. D. Bathgate Daniel J. Scott Thomas R. Weikl Guy N. L. Jameson Paul R. Gooley Unravelling the mechanism of neurotensin recognition by neurotensin receptor 1 Nature Communications |
| title | Unravelling the mechanism of neurotensin recognition by neurotensin receptor 1 |
| title_full | Unravelling the mechanism of neurotensin recognition by neurotensin receptor 1 |
| title_fullStr | Unravelling the mechanism of neurotensin recognition by neurotensin receptor 1 |
| title_full_unstemmed | Unravelling the mechanism of neurotensin recognition by neurotensin receptor 1 |
| title_short | Unravelling the mechanism of neurotensin recognition by neurotensin receptor 1 |
| title_sort | unravelling the mechanism of neurotensin recognition by neurotensin receptor 1 |
| url | https://doi.org/10.1038/s41467-023-44010-7 |
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