title: Photocurrent attenuation by a single polar-to-nonpolar point mutation of channelrhodopsin-2.
abstract: Channelrhodopsin-2 (ChR2), one of the algal light-gated cation channel rhodopsins, contains five peculiar glutamic acid residues in the N-terminal region corresponding to the second to third transmembrane helices. Here we made systematic mutations of these polar amino acid residues of ChR2 into nonpolar alanine, and evaluated their photocurrent properties. Amongst them, the photocurrent generated by the E97A mutation, ChR2(E97A), was much smaller than expected from its expression. The ChR2(E97A) photocurrent was similar to wild-type ChR2 in the kinetic profiles, the reversal potential and the dependency to the light power density. Our results suggest that the residue E97 is one of the molecular determinants involved in the ion flux regulation.
authors.affiliation: Department of Physiology and Pharmacology, Tohoku University Graduate School of Medicine, Sendai, 980-8575, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Sugiyama
authors.forename: Hongxia
authors.initials: H
authors.lastname: Wang
authors.forename: Takuya
authors.initials: T
authors.lastname: Hikima
authors.forename: Minami
authors.initials: M
authors.lastname: Sato
authors.forename: Jun
authors.initials: J
authors.lastname: Kuroda
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Takahashi
authors.forename: Toru
authors.initials: T
authors.lastname: Ishizuka
authors.forename: Hiromu
authors.initials: H
authors.lastname: Yawo
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D004594:Electrophysiology; D005453:Fluorescence; D006801:Humans; D015640:Ion Channel Gating / Q000528:radiation effects; D007700:Kinetics; D008027:Light; D008564:Membrane Potentials / Q000528:radiation effects*; D008969:Molecular Sequence Data; D017354:Point Mutation
chemical_list: D002352:Carrier Proteins; C523009:channelopsin-2, Chlamydomonas reinhardtii