title: pH dependence of Anabaena sensory rhodopsin: retinal isomer composition, rate of dark adaptation, and photochemistry.
abstract: Microbial rhodopsins are photoactive proteins, and their binding site can accommodate either all-trans or 13-cis retinal chromophore. The pH dependence of isomeric composition, dark-adaptation rate, and primary events of Anabaena sensory rhodopsin (ASR), a microbial rhodopsin discovered a decade ago, are presented. The main findings are: (a) Two pKa values of 6.5 and 4.0 assigned to two different protein residues are observed using spectroscopic titration experiments for both ground-state retinal isomers: all-trans, 15-anti (AT) and 13-cis, 15-syn (13C). The protonation states of these protein residues affect the absorption spectrum of the pigment and most probably the isomerization process of the retinal chromophore. An additional pKa value of 8.5 is observed only for 13C-ASR. (b) The isomeric composition of ASR is determined over a wide pH range and found to be almost pH-independent in the dark (>96% AT isomer) but highly pH-dependent in the light-adapted form. (c) The kinetics of dark adaptation is recorded over a wide pH range, showing that the thermal isomerization from 13C to AT retinal occurs much faster at high pH rather than under acidic conditions. (d) Primary photochemical events of ASR at pH 5 are recorded using VIS hyperspectral pump-probe spectroscopy with <100 fs resolution and compared with the previously recorded results at pH 7.5. For AT-ASR, these are shown to be almost pH-independent. However, photochemistry of 13C-ASR is pH-dependent and slowed down in acidic environments.
authors.affiliation: Department of Organic Chemistry, Weizmann Institute of Science , Rehovot 76100, Israel.
authors.forename: Rinat
authors.initials: R
authors.lastname: Rozin
authors.forename: Amir
authors.initials: A
authors.lastname: Wand
authors.forename: Kwang-Hwan
authors.initials: KH
authors.lastname: Jung
authors.forename: Sanford
authors.initials: S
authors.lastname: Ruhman
authors.forename: Mordechai
authors.initials: M
authors.lastname: Sheves
mesh_terms: D017033:Anabaena; D002851:Chromatography, High Pressure Liquid; D003624:Darkness; D004926:Escherichia coli; D006863:Hydrogen-Ion Concentration; D007536:Isomerism; D007700:Kinetics; D015394:Molecular Structure; D010775:Photic Stimulation; D055668:Photochemical Processes; D025603:Sensory Rhodopsins / Q000737:chemistry*; D013057:Spectrum Analysis; D013696:Temperature
chemical_list: D025603:Sensory Rhodopsins