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Title: The multidrug-resistance transporter MdfA from Escherichia coli: crystallization and X-ray diffraction analysis
Authors: Nagarathinam, Kumar
Jaenecke, Frank
Nakada-Nakura, Yoshiko
Hotta, Yunhon
Liu, Kehong
Iwata, So  kyouindb  KAKEN_id
Stubbs, Milton T.
Nomura, Norimichi  kyouindb  KAKEN_id
Tanabe, Mikio
Author's alias: 野村, 紀通
Keywords: MFS transporter
multidrug resistance
membrane protein
crystallization
antibody fragment
lipidic cubic phase
Issue Date: 1-Jul-2017
Publisher: International Union of Crystallography (IUCr)
Journal title: Acta Crystallographica Section F
Volume: 73
Start page: 423
End page: 430
Abstract: 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.
Rights: 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(Published Version): 10.1107/S2053230X17008500
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