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@ARTICLE{Bestsennaia:1024667,
      author       = {Bestsennaia, Ekaterina and Maslov, Ivan and Balandin, Taras
                      and Alekseev, Alexey and Yudenko, Anna and Abu Shamseye,
                      Assalla and Zabelskii, Dmitrii and Baumann, Arnd and
                      Catapano, Claudia and Karathanasis, Christos and Gordeliy,
                      Valentin and Heilemann, Mike and Gensch, Thomas and
                      Borshchevskiy, Valentin},
      title        = {{C}hannelrhodopsin‐2 {O}ligomerization in {C}ell
                      {M}embrane {R}evealed by {P}hoto‐{A}ctivated
                      {L}ocalization {M}icroscopy},
      journal      = {Angewandte Chemie},
      volume       = {136},
      number       = {11},
      issn         = {0932-2132},
      address      = {Weinheim},
      publisher    = {Wiley-VCH},
      reportid     = {FZJ-2024-02340},
      pages        = {e202307555},
      year         = {2024},
      note         = {We are thankful to Fedor Tsybrov for the help with
                      thepreparation of plasmids. V.B. acknowledges DAAD
                      YoungTalents Programme Line A. V.G. acknowledges his
                      HGFProfessorship. I.M. acknowledges FWO Research
                      FoundationFlanders (G0B9922N) and BOF UHasselt(BOF21BL11).
                      C.C., C.K. and M.H. gratefully acknowledgethe Deutsche
                      Forschungsgemeinschaft (grants CRC1507 andCRC807) for
                      financial support. The work was done in theframework of
                      CEA(IBS)–HGF(FZJ) STC 5.1 specific agreement.Open Access
                      funding enabled and organized byProjekt DEAL.},
      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/μm2.
                      Our findings are important for understanding the mechanistic
                      basis of ChR2 activity as well as for improving existing and
                      developing future optogenetic tools.},
      cin          = {IBI-1 / IBI-7},
      ddc          = {660},
      cid          = {I:(DE-Juel1)IBI-1-20200312 / I:(DE-Juel1)IBI-7-20200312},
      pnm          = {5241 - Molecular Information Processing in Cellular Systems
                      (POF4-524)},
      pid          = {G:(DE-HGF)POF4-5241},
      typ          = {PUB:(DE-HGF)16},
      doi          = {10.1002/ange.202307555},
      url          = {https://juser.fz-juelich.de/record/1024667},
}