TY  - JOUR
AU  - Bestsennaia, Ekaterina
AU  - Maslov, Ivan
AU  - Balandin, Taras
AU  - Alekseev, Alexey
AU  - Yudenko, Anna
AU  - Abu Shamseye, Assalla
AU  - Zabelskii, Dmitrii
AU  - Baumann, Arnd
AU  - Catapano, Claudia
AU  - Karathanasis, Christos
AU  - Gordeliy, Valentin
AU  - Heilemann, Mike
AU  - Gensch, Thomas
AU  - Borshchevskiy, Valentin
TI  - Channelrhodopsin‐2 Oligomerization in Cell Membrane Revealed by Photo‐Activated Localization Microscopy
JO  - Angewandte Chemie
VL  - 63
IS  - 11
SN  - 1433-7851
CY  - Weinheim
PB  - Wiley-VCH
M1  - FZJ-2024-01648
SP  - e202307555
PY  - 2024
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - 38226794
UR  - <Go to ISI:>//WOS:001155410200001
DO  - DOI:10.1002/anie.202307555
UR  - https://juser.fz-juelich.de/record/1023075
ER  -