Glutamate residue 90 in the predicted transmembrane domain 2 is crucial for cation flux through channelrhodopsin 2

Biochem Biophys Res Commun. 2011 Jul 15;410(4):737-43. doi: 10.1016/j.bbrc.2011.06.024. Epub 2011 Jun 12.


Channelrhodopsin 2 (ChR2) is a microbial-type rhodopsin with a putative heptahelical structure that binds all-trans-retinal. Blue light illumination of ChR2 activates an intrinsic leak channel conductive for cations. Sequence comparison of ChR2 with the related ChR1 protein revealed a cluster of charged amino acids within the predicted transmembrane domain 2 (TM2), which includes glutamates E90, E97 and E101. Charge inversion substitutions of these residues significantly altered ChR2 function as revealed by two-electrode voltage-clamp recordings of light-induced currents from Xenopus laevis oocytes expressing the respective mutant proteins. Specifically, replacement of E90 by lysine or alanine resulted in differential effects on H(+)- and Na(+)-mediated currents. Our results are consistent with this glutamate side chain within the proposed TM2 contributing to ion flux through and the cation selectivity of ChR2.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cations / metabolism
  • Chlamydomonas reinhardtii / genetics
  • Chlamydomonas reinhardtii / metabolism
  • Glutamic Acid / chemistry
  • Glutamic Acid / genetics
  • Hydrogen-Ion Concentration
  • Ion Transport
  • Molecular Sequence Data
  • Protein Structure, Secondary
  • Protein Structure, Tertiary / genetics
  • Rhodopsin / chemistry
  • Rhodopsin / genetics
  • Rhodopsin / metabolism*
  • Xenopus laevis


  • Cations
  • Glutamic Acid
  • Rhodopsin