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Expression, purification, crystallization and preliminary X-ray analysis of the DNA-binding domain of Rhodobacter capsulatus MopB.

ABSTRACT: The LysR-type regulator MopB represses transcription of several target genes (including the nitrogen-fixation gene anfA) in Rhodobacter capsulatus at high molybdenum concentrations. In this study, the isolated DNA-binding domain of MopB (MopBHTH) was overexpressed in Escherichia coli. Purified MopBHTH bound the anfA promoter as shown by DNA mobility-shift assays, demonstrating the function of the isolated regulator domain. MopBHTH was crystallized using the sitting-drop vapour-diffusion method in the presence of 0.2?M lithium sulfate, 0.1?M phosphate/citrate pH 4.2, 20%(w/v) PEG 1000 at 291?K. The crystal belonged to space group P3(1)21 or P3(2)21, with unit-cell parameters a=b=61.84, c=139.64?Å, ?=?=90, ?=120°, and diffracted to 3.3?Å resolution at a synchrotron source.

SUBMITTER: Muller A

PROVIDER: S-EPMC3053167 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature

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Expression, purification, crystallization and preliminary X-ray analysis of the DNA-binding domain of Rhodobacter capsulatus MopB.

Müller Alexandra A Schlicker Christine C Fehringer Maria M Masepohl Bernd B Hofmann Eckhard E

Acta crystallographica. Section F, Structural biology and crystallization communications 20110225 Pt 3

The LysR-type regulator MopB represses transcription of several target genes (including the nitrogen-fixation gene anfA) in Rhodobacter capsulatus at high molybdenum concentrations. In this study, the isolated DNA-binding domain of MopB (MopBHTH) was overexpressed in Escherichia coli. Purified MopBHTH bound the anfA promoter as shown by DNA mobility-shift assays, demonstrating the function of the isolated regulator domain. MopBHTH was crystallized using the sitting-drop vapour-diffusion method i ...[more]

PMID: 21393847

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