Biochemistry (Moscow)

, Volume 80, Issue 4, pp 449–454 | Cite as

Proteorhodopsin from Dokdonia sp. PRO95 is a light-driven Na+-pump

  • Y. V. Bertsova
  • A. V. Bogachev
  • V. P. Skulachev
Article

Abstract

The gene encoding proteorhodopsin AEX55013 from Dokdonia sp. PRO95 was cloned and expressed in Escherichia coli cells. Illumination of the proteorhodopsin-producing E. coli cells in Na+-containing media resulted in alkalinization of the media. This response was accelerated by uncoupler CCCP and inhibited by penetrating anion SCN. Illumination of the cells in a sodium-free medium (made by substituting Na+ with K+) resulted in SCN-stimulated and CCCP-sensitive acidification of the medium. Illumination of the proteorhodopsin-containing E. coli cells caused CCCP-resistant transmembrane sodium export from these cells. We conclude that the proteorhodopsin from the marine flavobacterium Dokdonia sp. PRO95 is a primary light-driven Na+-pump. A high level of the heterologous production in E. coli cells as well as stability and purity of the isolated protein makes this proteorhodopsin an attractive model for studying the mechanism of active sodium transmembrane translocation.

Key words

Na+-translocating proteorhodopsin transmembrane sodium transport flavobacteria 

Abbreviations

CCCP

carbonyl cyanide m-chlorophenylhydrazone

DDM

n-dodecyl β-D-maltoside

R-E. coli

E. coli BL21/pRhod_10.1 cells induced for proteorhodopsin synthesis in the presence of retinal

SDS-PAGE

sodium dodecyl sulfate polyacrylamide gel electrophoresis

ΔpH

transmembrane pH difference

\(\Delta \bar \mu _{H^ + } \) and \(\Delta \bar \mu _{Na^ + } \)

transmembrane differences in H+ and Na+ electrochemical potentials, respectively

Δψ

transmembrane difference in electric potentials

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Copyright information

© Pleiades Publishing, Ltd. 2015

Authors and Affiliations

  • Y. V. Bertsova
    • 1
  • A. V. Bogachev
    • 1
  • V. P. Skulachev
    • 1
  1. 1.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia

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