Retinal-based anion pump from the cyanobacterium Tolypothrix campylonemoides

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Abstract

In this work, a functional characterization of TcaR rhodopsin from the cyanobacterium Tolypothrix campylonemoides was obtained. Analysis of the amino acid sequence of TcaR revealed that this protein possesses a TSD motif that differs by only one amino acid from the motif of the known halorodopsin chloride pump TSA. The TcaR protein was expressed in E. coli, purified and incorporated into proteoliposomes and nanodiscs. Functional activity was measured by electric current generation through the planar bilayer lipid membranes (BLMs) with proteoliposomes adsorbed on one membrane surface, as well as by fluorescence using voltage-dependent dye oxonol VI. We have shown that TcaR rhodopsin functions as a powerful anion pump. Our results show that the novel microbial anion transporter TcaR deserves deeper study and may be of interest both for fundamental studies of membrane proteins and as a tool for optogenetics.

About the authors

T. I Rokitskaya

Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University

Email: rokitskaya@belozersky.msu.ru
119991 Moscow, Russia

A. A Alekseev

Moscow Institute of Physics and Technology

141701 Dolgoprudny, Moscow Region, Russia

F. M Tsybrov

Moscow Institute of Physics and Technology

141701 Dolgoprudny, Moscow Region, Russia

S. M Bukhalovich

Moscow Institute of Physics and Technology

141701 Dolgoprudny, Moscow Region, Russia

Y. N Antonenko

Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University

Email: antonen@belozersky.msu.ru
119991 Moscow, Russia

V. I Gordeliy

Institut de Biologie Structurale (IBS), Universit� Grenoble Alpes, CEA, CNRS

Email: valentin.gordeliy@gmail.com
Grenoble, France

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