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タイトル: | The multidrug-resistance transporter MdfA from Escherichia coli: crystallization and X-ray diffraction analysis |
著者: | Nagarathinam, Kumar Jaenecke, Frank Nakada-Nakura, Yoshiko Hotta, Yunhon Liu, Kehong Iwata, So Stubbs, Milton T. Nomura, Norimichi https://orcid.org/0000-0002-6330-2239 (unconfirmed) Tanabe, Mikio |
著者名の別形: | 野村, 紀通 |
キーワード: | MFS transporter multidrug resistance membrane protein crystallization antibody fragment lipidic cubic phase |
発行日: | 1-Jul-2017 |
出版者: | International Union of Crystallography (IUCr) |
誌名: | Acta Crystallographica Section F |
巻: | 73 |
開始ページ: | 423 |
終了ページ: | 430 |
抄録: | The active efflux of antibiotics by multidrug-resistance (MDR) transporters is a major pathway of drug resistance and complicates the clinical treatment of bacterial infections. MdfA is a member of the major facilitator superfamily (MFS) from Escherichia coli and provides resistance to a wide variety of dissimilar toxic compounds, including neutral, cationic and zwitterionic substances. The 12-transmembrane-helix MdfA was expressed as a GFP-octahistidine fusion protein with a TEV protease cleavage site. Following tag removal, MdfA was purified using two chromatographic steps, complexed with a Fab fragment and further purified using size-exclusion chromatography. MdfA and MdfA–Fab complexes were subjected to both vapour-diffusion and lipidic cubic phase (LCP) crystallization techniques. Vapour-diffusion-grown crystals were of type II, with poor diffraction behaviour and weak crystal contacts. LCP lipid screening resulted in type I crystals that diffracted to 3.4 Å resolution and belonged to the hexagonal space group P6[1]22. |
著作権等: | Copyright © International Union of Crystallography Author(s) of this paper may load this reprint on their own web site or institutional repository provided that this cover page is retained. Republication of this article or its storage in electronic databases other than as specified above is not permitted without prior permission in writing from the IUCr. |
URI: | http://hdl.handle.net/2433/226310 |
DOI(出版社版): | 10.1107/S2053230X17008500 |
PubMed ID: | 28695852 |
出現コレクション: | 学術雑誌掲載論文等 |
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