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

Kikukawa, Takashi; Miyauchi, Seiji; Araiso, Tsunehisa; Kamo, Naoki; Nara, Toshifumi

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

Kikukawa, Takashi; Miyauchi, Seiji; Araiso, Tsunehisa et al.

Biochemical and Biophysical Research Communications, 2007, 358(4), 1071-1075

Permalink : https://hdl.handle.net/2115/28010

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アクセス権	open access
URI	https://hdl.handle.net/2115/28010
タイトル	Anti-parallel membrane topology of two components of EbrAB, a multidrug transporter.
著者	Kikukawa, Takashi ; Miyauchi, Seiji ; Araiso, Tsunehisa ; Kamo, Naoki ; Nara, Toshifumi
e-Rad_Researcher 20281842 著者名 Kikukawa, Takashi
著者名 Miyauchi, Seiji
著者名 Araiso, Tsunehisa
著者名 Kamo, Naoki
著者名 Nara, Toshifumi
言語	英語
キーワード	Efflux transporter
	EmrE
	Small multidrug resistance (SMR)
	Dual topology configuration
	Fluorescein-5-maleimide (NEM-fluorescein)
	Ethidium efflux
発行日	2007-07-13
出版者	Elsevier Inc.
収録物名	Biochemical and Biophysical Research Communications
巻(号)	358(4)
ページ	1071-1075
抄録	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.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
PISSN	0006-291X
EISSN	1090-2104
関連情報	PMID 17517373
関連情報	URI http://www.sciencedirect.com/science/journal/0006291X
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.bbrc.2007.05.032