Journal Article

FZJ-2024-01648

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Channelrhodopsin‐2 Oligomerization in Cell Membrane Revealed by Photo‐Activated Localization Microscopy

Bestsennaia, E. ; Maslov, I. ; Balandin, T.FZJ* ; Alekseev, A. ; Yudenko, A. ; Abu Shamseye, A.FZJ* ; Zabelskii, D. ; Baumann, A. ; Catapano, C. ; Karathanasis, C. ; Gordeliy, V.FZJ* ; Heilemann, M. ; Gensch, T. (Corresponding author)FZJ* ; Borshchevskiy, V.

2024
Wiley-VCH Weinheim

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Please use a persistent id in citations: doi:10.1002/anie.202307555  doi:10.34734/FZJ-2024-01648

Abstract: Microbialrhodopsinsare retinalmembraneproteinsthat founda broadapplicationin optogenetics.Theoligomericstate of rhodopsinsis importantfor their functionalityand stability.Of particularinterestis the oligomericstate in the cellularnativemembraneenvironment.Fluorescencemicroscopyprovidespowerfultools to determinetheoligomericstate of membraneproteinsdirectlyin cells. Amongthese methodsis quantitativephotoactivatedlocalizationmicroscopy(qPALM)allowingthe investigationof molecularorganizationat the level of singleproteinclusters.Here,we applyqPALMto investigatethe oligomericstate of the first and most used optogenetictool Channelrhodopsin-2(ChR2)in the plasmamembraneof eukaryoticcells. ChR2appearedpredominantlyas a dimerin the cell membraneanddid not form higheroligomers.The disulfidebondsbetweenCys34and Cys36of adjacentChR2monomerswere notrequiredfor dimerformationand mutationsdisruptingthese bondsresultedin only partialmonomerizationof ChR2.The monomericfractionincreasedwhenthe total concentrationof mutantChR2in the membranewas low. Thedissociationconstantwas estimatedfor this partiallymonomerizedmutantChR2as 2.2�0.9 proteins/μm2. Our findingsare importantfor understandingthe mechanisticbasis of ChR2activityas well as for improvingexistingand developingfutureoptogenetictools.

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Contributing Institute(s):

1. Molekular- und Zellphysiologie (IBI-1)
2. Strukturbiochemie (IBI-7)

Research Program(s):

1. 5241 - Molecular Information Processing in Cellular Systems (POF4-524) (POF4-524)

Appears in the scientific report 2024

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Database coverage:
; ; ; Chemical Reactions ; Clarivate Analytics Master Journal List ; Current Contents - Life Sciences ; Current Contents - Physical, Chemical and Earth Sciences ; DEAL Wiley ; Ebsco Academic Search ; Essential Science Indicators ; IF >= 15 ; Index Chemicus ; JCR ; Nationallizenz ; SCOPUS ; Science Citation Index Expanded ; Web of Science Core Collection

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