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

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Title: Expression of salinarum halorhodopsin in Escherichia coli cells : Solubilization in the presence of retinal yields the natural state
Authors: Yamashita, Yasutaka Browse this author
Kikukawa, Takashi Browse this author →KAKEN DB
Tsukamoto, Takashi Browse this author
Kamiya, Masakatsu Browse this author
Aizawa, Tomoyasu Browse this author
Kawano, Keiichi Browse this author
Miyauchi, Seiji Browse this author
Kamo, Naoki Browse this author
Demura, Makoto Browse this author
Keywords: Archaeal rhodopsin
Halorhodopsin
Light-driven chloride pump
Photocycle
Issue Date: Dec-2011
Publisher: Elsevier B.V.
Journal Title: Biochimica et Biophysica Acta (BBA) : Biomembranes
Volume: 1808
Issue: 12
Start Page: 2905
End Page: 2912
Publisher DOI: 10.1016/j.bbamem.2011.08.035
PMID: 21925140
Abstract: Salinarum halorhodopsin (HsHR), a light-driven chloride ion pump of haloarchaeon Halobacterium salinarum, was heterologously expressed in Escherichia coil. 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.
Type: article (author version)
URI: http://hdl.handle.net/2115/48254
Appears in Collections:生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 菊川 峰志

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