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Title: | Sub-millisecond conformational dynamics of the A(2A) adenosine receptor revealed by single-molecule FRET | Authors: | MASLOV, Ivan Volkov, Oleksandr Khorn, Polina Orekhov, Philipp Gusach, Anastasiia Kuzmichev, Pavel Gerasimov, Andrey Luginina, Aleksandra Coucke, Quinten Bogorodskiy, Andrey Gordeliy, Valentin Wanninger, Simon Barth, Anders Mishin, Alexey Hofkens, Johan Cherezov, Vadim Gensch, Thomas HENDRIX, Jelle Borshchevskiy, Valentin |
Issue Date: | 2023 | Publisher: | NATURE PORTFOLIO | Source: | Communications Biology, 6 (1) (Art N° 362) | Abstract: | Single-molecule FRET experiments with human A2A adenosine receptor in its apo and agonist-bound states in lipid nanodiscs provide insights into its conformational dynamics and activation. The complex pharmacology of G-protein-coupled receptors (GPCRs) is defined by their multi-state conformational dynamics. Single-molecule Forster Resonance Energy Transfer (smFRET) is well suited to quantify dynamics for individual protein molecules; however, its application to GPCRs is challenging. Therefore, smFRET has been limited to studies of inter-receptor interactions in cellular membranes and receptors in detergent environments. Here, we performed smFRET experiments on functionally active human A(2A) adenosine receptor (A(2A)AR) molecules embedded in freely diffusing lipid nanodiscs to study their intramolecular conformational dynamics. We propose a dynamic model of A(2A)AR activation that involves a slow (>2 ms) exchange between the active-like and inactive-like conformations in both apo and antagonist-bound A(2A)AR, explaining the receptor's constitutive activity. For the agonist-bound A(2A)AR, we detected faster (390 +/- 80 mu s) ligand efficacy-dependent dynamics. Our work establishes a general smFRET platform for GPCR investigations that can potentially be used for drug screening and/or mechanism-of-action studies. | Notes: | Borshchevskiy, V (corresponding author), Moscow Inst Phys & Technol, Res Ctr Mol Mech Aging & Age Related Dis, Dolgoprudnyi, Moscow Region, Russia.; Hendrix, J (corresponding author), Hasselt Univ, Biomed Res Inst, Adv Opt Microscopy Ctr, Dynam Bioimaging Lab,Agoralaan C BIOMED, Diepenbeek, Belgium.; Hendrix, J (corresponding author), Katholieke Univ Leuven, Dept Chem, Div Mol Imaging & Photon, Lab Photochem & Spect, Leuven, Belgium.; Borshchevskiy, V (corresponding author), Joint Inst Nucl Res, Dubna, Russia. jelle.hendrix@uhasselt.be; borshchevskiy.vi@phystech.edu |
Keywords: | Humans;Molecular Conformation;Cell Membrane;Proteins;Fluorescence Resonance Energy Transfer;Receptor, Adenosine A2A | Document URI: | http://hdl.handle.net/1942/40052 | e-ISSN: | 2399-3642 | DOI: | 10.1038/s42003-023-04727-z | ISI #: | 000962868600002 | Rights: | The Author(s) 2023. 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/. | Category: | A1 | Type: | Journal Contribution |
Appears in Collections: | Research publications |
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