title: Redox potentials of the oriented film of the wild-type, the E194Q-, E204Q- and D96N-mutated bacteriorhodopsins.
abstract: The redox potentials of the oriented films of the wild-type, the E194Q-, E204Q- and D96N-mutated bacteriorhodopsins (bR), prepared by adsorbing purple membrane (PM) sheets or its mutant on a Pt electrode, have been examined. The redox potentials (V) of the wild-type bR were -470 mV for the 13-cis configuration of the retinal Shiff base in bR and -757 mV for the all-trans configuration in H(2)O, and -433 mV for the 13-cis configuration and -742 mV for the all-trans configuration in D(2)O. The solvent isotope effect (DeltaV=V(D(2)O)-V(H(2)O)), which shifts the redox potential to a higher value, originates from the cooperative rearrangements of the extensively hydrogen-bonded water molecules around the protonated C=N part in the retinal Schiff base. The redox potential of bR was much higher for the 13-cis configuration than that for the all-trans configuration. The redox potentials for the E194Q mutant in the extracellular region were -507 mV for the 13-cis configuration and -788 mV for the all-trans configuration; and for the E204Q mutant they were -491 mV for the 13-cis configuration and -769 mV for the all-trans configuration. Replacement of the Glu(194) or Glu(204) residues by Gln weakened the electron withdrawing interaction to the protonated C=N bond in the retinal Schiff base. The E204 residue is less linked with the hydrogen-bonded network of the proton release pathway compared with E194. The redox potentials of the D96N mutant in the cytoplasmic region were -471 mV for the 13-cis configuration and -760 mV for the all-trans configuration which were virtually the same as those of the wild-type bR, indicating that the D to N point mutation of the 96 residue had no influence on the interaction between the D96 residue and the C=N part in the Schiff base under the light-adapted condition. The results suggest that the redox potential of bR is closely correlated to the hydrogen-bonded network spanning from the retinal Schiff base to the extracellular surface of bR in the proton transfer pathway.
authors.affiliation: Faculty of Pharmaceutical Sciences, Tokushima University, Tokushima, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Ueno
authors.forename: Akira
authors.initials: A
authors.lastname: Shibata
authors.forename: Ayako
authors.initials: A
authors.lastname: Yorimitsu
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Baba
authors.forename: Naoki
authors.initials: N
authors.lastname: Kamo
mesh_terms: D001436:Bacteriorhodopsins / Q000235:genetics / Q000502:physiology*; D006217:Halobacterium / Q000502:physiology; D009154:Mutation; D010084:Oxidation-Reduction; D012997:Solvents
chemical_list: D012997:Solvents; D001436:Bacteriorhodopsins