Please use this identifier to cite or link to this item:
http://hdl.handle.net/1942/42484
Title: | Channelrhodopsin-2 Oligomerization in Cell Membrane Revealed by Photo-Activated Localization Microscopy | Authors: | Bestsennaia, Ekaterina MASLOV, Ivan Balandin, Taras Alekseev, Alexey Yudenko, Anna Abu Shamseye, Assalla Zabelskii, Dmitrii Baumann, Arnd Catapano, Claudia Karathanasis, Christos Gordeliy, Valentin Heilemann, Mike Gensch, Thomas Borshchevskiy, Valentin |
Issue Date: | 2024 | Publisher: | WILEY-V C H VERLAG GMBH | Source: | ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, | Abstract: | Microbial rhodopsins are retinal membrane proteins that found a broad application in optogenetics. The oligomeric state of rhodopsins is important for their functionality and stability. Of particular interest is the oligomeric state in the cellular native membrane environment. Fluorescence microscopy provides powerful tools to determine the oligomeric state of membrane proteins directly in cells. Among these methods is quantitative photoactivated localization microscopy (qPALM) allowing the investigation of molecular organization at the level of single protein clusters. Here, we apply qPALM to investigate the oligomeric state of the first and most used optogenetic tool Channelrhodopsin-2 (ChR2) in the plasma membrane of eukaryotic cells. ChR2 appeared predominantly as a dimer in the cell membrane and did not form higher oligomers. The disulfide bonds between Cys34 and Cys36 of adjacent ChR2 monomers were not required for dimer formation and mutations disrupting these bonds resulted in only partial monomerization of ChR2. The monomeric fraction increased when the total concentration of mutant ChR2 in the membrane was low. The dissociation constant was estimated for this partially monomerized mutant ChR2 as 2.2 +/- 0.9 proteins/mu m2. Our findings are important for understanding the mechanistic basis of ChR2 activity as well as for improving existing and developing future optogenetic tools. Insights from super-resolution microscopy: Channelrhodopsin-2 (ChR2), a widely used optogenetic tool, forms dimers, not higher oligomers, in human cell membranes. Disruption of inter-protein disulfide bonds leads to partial ChR2 monomerization, particularly in cells with lower ChR2 densities. This study enhances the understanding of ChR2 oligomerization and assists optogenetic tool design. image | Notes: | Gensch, T (corresponding author), Forschungszentrum Julich, Inst Biol Informat Proc 1, Mol & Cellular Physiol IBI-1, D-52428 Julich, Germany.; Borshchevskiy, V (corresponding author), Forschungszentrum Julich, Inst Biol Informat Proc 7, Struct Biochem IBI-7, D-52428 Julich, Germany. t.gensch@fz-juelich.de; v.borshchevskiy@fz-juelich.de |
Keywords: | channelrhodopsin-2;membrane proteins;oligomerization;optogenetic tools;super-resolution microscopy | Document URI: | http://hdl.handle.net/1942/42484 | ISSN: | 1433-7851 | e-ISSN: | 1521-3773 | DOI: | 10.1002/anie.202307555 | ISI #: | 001155410200001 | Rights: | 2024The Authors. AngewandteChemieInternationalEditionpublishedby Wiley-VCHGmbH.This is an openaccessarticleunderthe termsof the CreativeCommonsAttributionLicense,whichpermitsuse, distributionand reproductionin any medium,providedthe originalworkis properlycited | Category: | A1 | Type: | Journal Contribution |
Appears in Collections: | Research publications |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
Angew Chem Int Ed - 2024 - Bestsennaia - Channelrhodopsin‐2.pdf | Early view | 4.5 MB | Adobe PDF | View/Open |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.