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Anti-parallel membrane topology of two components of EbrAB, a multidrug transporter.

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Title: Anti-parallel membrane topology of two components of EbrAB, a multidrug transporter.
Authors: Kikukawa, Takashi Browse this author →KAKEN DB
Miyauchi, Seiji Browse this author
Araiso, Tsunehisa Browse this author
Kamo, Naoki Browse this author
Nara, Toshifumi Browse this author
Keywords: Efflux transporter
EmrE
Small multidrug resistance (SMR)
Dual topology configuration
Fluorescein-5-maleimide (NEM-fluorescein)
Ethidium efflux
Issue Date: 13-Jul-2007
Publisher: Elsevier Inc.
Journal Title: Biochemical and Biophysical Research Communications
Volume: 358
Issue: 4
Start Page: 1071
End Page: 1075
Publisher DOI: 10.1016/j.bbrc.2007.05.032
PMID: 17517373
Abstract: EbrAB is a multidrug-resistance transporter in Bacillus subtilis that belongs to the small multidrug resistance, and requires two polypeptides of both EbrA and EbrB, implying that it functions in the hetero-dimeric state. In this study, we investigated the transmembrane topologies of EbrA and EbrB. Various single-cysteine mutants were expressed in Escherichia coli cells, and the efflux activity was measured. Only mutants having a high activity were used for the topology experiments. The reactivity of a membrane impermeable NEM-fluorescein against the single cysteine of these fully functional mutants was examined when this reactive fluorophore was applied either from the outside or both sides of the cell membrane or in the denatured state. The results clearly showed that EbrA and EbrB have the opposite orientation within the membrane or an anti-parallel configuration.
Relation: http://www.sciencedirect.com/science/journal/0006291X
Type: article (author version)
URI: http://hdl.handle.net/2115/28010
Appears in Collections:創成研究機構 (Creative Research Institution) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 菊川 峰志

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