Ion channels undergo major conformational changes that lead to channel opening and ion conductance. Deciphering these structure-function relationships is paramount to understanding channel physiology and pathophysiology. Cryo-electron microscopy, crystallography and computer modelling provide atomic-scale snapshots of channel conformations in non-cellular environments but lack dynamic information that can be linked to functional results. Biophysical techniques such as electrophysiology, on the other hand, provide functional data with no structural information of the processes involved. Fluorescence spectroscopy techniques help bridge this gap in simultaneously obtaining structure-function correlates. These include voltage-clamp fluorometry, Förster resonance energy transfer, ligand binding assays, single molecule fluorescence and their variations. These techniques can be employed to unearth several features of ion channel behaviour. For instance, they provide real time information on local and global rearrangements that are inherent to channel properties. They also lend insights in trafficking, expression, and assembly of ion channels on the membrane surface. These methods have the advantage that they can be carried out in either native or heterologous systems. In this review, we briefly explain the principles of fluorescence and how these have been translated to study ion channel function. We also report several recent advances in fluorescence spectroscopy that has helped address and improve our understanding of the biophysical behaviours of different ion channel families.
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October 2022
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Protein crowding induces membrane curvatures through an entropic mechanism. Crowding of asymmetric proteins results in an asymmetric lateral pressure across the membrane which can be used by cells in a number of biological processes involving membrane remodeling. For further information, see the review in this issue by Ruhoff and colleagues (pages 1257–1267). Image provided by Poul Martin Bendix.
Review Article|
October 25 2022
Characterising ion channel structure and dynamics using fluorescence spectroscopy techniques
Shreyas Bhat
;
Shreyas Bhat
1Department of Physics, Université de Montréal, Montréal, QC, Canada
2Department of Pharmacology and Physiology, Université de Montréal, Montréal, QC, Canada
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Rikard Blunck
1Department of Physics, Université de Montréal, Montréal, QC, Canada
2Department of Pharmacology and Physiology, Université de Montréal, Montréal, QC, Canada
3Center for Interdisciplinary Research on Brain and Learning (CIRCA), Université de Montréal, Montréal, QC, Canada
Correspondence: Rikard Blunck (rikard.blunck@umontreal.ca)
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Publisher: Portland Press Ltd
Received:
July 31 2022
Revision Received:
September 21 2022
Accepted:
October 04 2022
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2022 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society
2022
Biochem Soc Trans (2022) 50 (5): 1427–1445.
Article history
Received:
July 31 2022
Revision Received:
September 21 2022
Accepted:
October 04 2022
Citation
Shreyas Bhat, Rikard Blunck; Characterising ion channel structure and dynamics using fluorescence spectroscopy techniques. Biochem Soc Trans 31 October 2022; 50 (5): 1427–1445. doi: https://doi.org/10.1042/BST20220605
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