His57 controls the efficiency of ESR, a light-driven proton pump from Exiguobacterium sibiricum at low and high pH

Siletsky, S.A.; Lukashev, E.P.; Mamedov, M.D.; Borisov, V.B.; Balashov, S.P.; Dolgikh, D.A.; Rubin, A.B.; Kirpichnikov, M.P.; Petrovskaya, L.E.

Авторы: Siletsky Sergey A., Lukashev Evgeniy P., Mamedov Mahir D., Borisov Vitaliy B., Balashov Sergei P., Dolgikh Dmitriy A., Rubin Andrei B., Kirpichnikov Mikhail P., Petrovskaya Lada E.
Журнал: Biochimica et Biophysica Acta - Bioenergetics
Том: 1862
Номер: 1
Год издания: 2021
Издательство: Elsevier BV
Местоположение издательства: Netherlands
Номер статьи: 148328
DOI: 10.1016/j.bbabio.2020.148328
Аннотация: ESR, a light-driven proton pump from Exiguobacterium sibiricum, contains a lysine residue (Lys96) in the proton donor site. Substitution of Lys96 with a nonionizable residue greatly slows reprotonation of the retinal Schiff base. The recent study of electrogenicity of the K96A mutant revealed that overall efficiency of proton transport is decreased in the mutant due to back reactions (Siletsky et al., BBA, 2019). Similar to members of the proteorhodopsin and xanthorhodopsin families, in ESR the primary proton acceptor from the Schiff base, Asp85, closely interacts with His57. To examine the role of His57 in the efficiency of proton translocation by ESR, we studied the effects of H57N and H57N/K96A mutations on the pH dependence of light-induced pH changes in suspensions of Escherichia coli cells, kinetics of absorption changes and electrogenic proton transfer reactions during the photocycle. We found that at low pH (<5) the proton pumping efficiency of the H57N mutant in E. coli cells and its electrogenic efficiency in proteoliposomes is substantially higher than in the WT, suggesting that interaction of His57 with Asp85 sets the low pH limit for H+ pumping in ESR. The electrogenic components that correspond to proton uptake were strongly accelerated at low pH in the mutant indicating that Lys96 functions as a very efficient proton donor at low pH. In the H57N/K96A mutant, a higher H+ pumping efficiency compared with K96A was observed especially at high pH, apparently from eliminating back reactions between Asp85 and the Schiff base by the H57N mutation.
Добавил в систему: Борисов Виталий Борисович

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His57 controls the efficiency of ESR, a light-driven proton pump from Exiguobacterium sibiricum at low and high pHстатья

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