期刊
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS
卷 73, 期 -, 页码 423-430出版社
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S2053230X17008500
关键词
MFS transporter; multidrug resistance; membrane protein; crystallization; antibody fragment; lipidic cubic phase
资金
- Bundesministerium fur Bildung und Forschung (BMBF) (ZIK program) [FKZ 03Z2HN21]
- ERDF [1241090001]
- Platform Project for Support in Drug Discovery and Life Science Research (Platform for Drug Discovery, Informatics and Structural Life Science) from the Japan Agency for Medical Research and Development (AMED)
- ERATO Human Receptor Crystallography Project of the Japan Science and Technology Agency (JST)
- Strategic Basic Research Program, JST
- Research Acceleration Program of the JST
- Targeted Proteins Research Program of the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan
- MEXT [22570114]
- BioStruct-X [5450, 8015]
- Grants-in-Aid for Scientific Research [15K06968, 22570114] Funding Source: KAKEN
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 angstrom resolution and belonged to the hexagonal space group P6(1)22.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据