Expression of salinarum halorhodopsin in Escherichia coli cells: Solubilization in the presence of retinal yields the natural state

Yasutaka Yamashita, Takashi Kikukawa, Takashi Tsukamoto, Masakatsu Kamiya, Tomoyasu Aizawa, Keiichi Kawano, Seiji Miyauchi, Naoki Kamo, Makoto Demura

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18 Citations (Scopus)

Abstract

Salinarum halorhodopsin (HsHR), a light-driven chloride ion pump of haloarchaeon Halobacterium salinarum, was heterologously expressed in Escherichia coli. The expressed HsHR had no color in the E. coli membrane, but turned purple after solubilization in the presence of all-trans retinal. This colored HsHR was purified by Ni-chelate chromatography in a yield of 3-4 mg per liter culture. The purified HsHR showed a distinct chloride pumping activity by incorporation into the liposomes, and showed even in the detergent-solubilized state, its typical behaviors in both the unphotolyzed and photolyzed states. Upon solubilization, HsHR expressed in the E. coli membrane attains the proper folding and a trimeric assembly comparable to those in the native membranes.

Original languageEnglish
Pages (from-to)2905-2912
Number of pages8
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1808
Issue number12
DOIs
Publication statusPublished - Dec 1 2011

Keywords

  • Archaeal rhodopsin
  • Halorhodopsin
  • Light-driven chloride pump
  • Photocycle

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

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    Yamashita, Y., Kikukawa, T., Tsukamoto, T., Kamiya, M., Aizawa, T., Kawano, K., Miyauchi, S., Kamo, N., & Demura, M. (2011). Expression of salinarum halorhodopsin in Escherichia coli cells: Solubilization in the presence of retinal yields the natural state. Biochimica et Biophysica Acta - Biomembranes, 1808(12), 2905-2912. https://doi.org/10.1016/j.bbamem.2011.08.035