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Medicinal-Pharmaceutical Chemistry Commons

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Physics and Astronomy Publications

2006

Articles 1 - 2 of 2

Full-Text Articles in Medicinal-Pharmaceutical Chemistry

Cloning, Expression, Purification, Crystallization And Preliminary X-Ray Diffraction Analysis Of The Regulator Acrr From Escherichia Coli, Ming Li, Xi Qiu, Chih-Chia Su, Feng Long, Ruoyu Gu, Gerry Mcdermott, Edward Yu Jan 2006

Cloning, Expression, Purification, Crystallization And Preliminary X-Ray Diffraction Analysis Of The Regulator Acrr From Escherichia Coli, Ming Li, Xi Qiu, Chih-Chia Su, Feng Long, Ruoyu Gu, Gerry Mcdermott, Edward Yu

Physics and Astronomy Publications

This paper describes the cloning, expression, purification and preliminary X-ray data analysis of the AcrR regulatory protein. The Escherichia coli AcrR is a member of the TetR family of transcriptional regulators. It regulates the expression of the AcrAB multidrug transporter. Recombinant AcrR with a 6×His tag at the C-terminus was expressed in E. coli and purified by metal-affinity chromatography. The protein was crystallized using hanging-drop vapor diffusion. X-ray diffraction data were collected from cryocooled crystals at a synchrotron light source. The best crystal diffracted to 2.5 Å. The space group was determined to be P32, with ...


Conformation Of The Acrb Multidrug Efflux Pump In Mutants Of The Putative Proton Relay Pathway, Chih-Chia Su, Ming Li, Ruoyu Gu, Yumiko Takatsuka, Gerry Mcdermott, Hiroshi Nikaido, Edward Yu Jan 2006

Conformation Of The Acrb Multidrug Efflux Pump In Mutants Of The Putative Proton Relay Pathway, Chih-Chia Su, Ming Li, Ruoyu Gu, Yumiko Takatsuka, Gerry Mcdermott, Hiroshi Nikaido, Edward Yu

Physics and Astronomy Publications

We previously reported the X-ray structures of wild-type Escherichia coli AcrB, a proton motive force-dependent multidrug efflux pump, and its N109A mutant. These structures presumably reflect the resting state of AcrB, which can bind drugs. After ligand binding, a proton may bind to an acidic residue(s) in the transmembrane domain, i.e., Asp407 or Asp408, within the putative network of electrostatically interacting residues, which also include Lys940 and Thr978, and this may initiate a series of conformational changes that result in drug expulsion. Herein we report the X-ray structures of four AcrB mutants, the D407A, D408A, K940A, and T978A ...