Expression, purification, crystallization and preliminary crystallographic analysis of the mouse transcription factor MafB in complex with its DNA-recognition motif Cmare
The MafB transcription factor (residues 211–305) has been crystallized in complex with the 21 bp Cmare DNA-binding site. Native and mercury-derivatized data collection and preliminary analyses are reported. The MafB transcription factor (residues 211–305) has been overexpressed in and purified from Escherichia coli. A protein–DNA complex between the MafB homodimer and the 21 bp Maf-recognition sequence known as Cmare has been successfully reconstituted in vitro and subsequently crystallized. The diffraction properties of the protein–DNA complex crystals were improved using a combination of protein-construct boundary optimization and targeted mutagenesis to promote crystal lattice stability. Both native and mercury-derivatized crystals have been prepared using these optimized conditions. The crystals belong to space group P4{sub 1}2{sub 1}2 or P4{sub 3}2{sub 1}2, with unit-cell parameters a = b = 94.8, c = 197.9 Å. An anomalous difference Patterson map computed using data collected from crystals grown in the presence of HgCl{sub 2} reveals four peaks. This corresponds to two copies of the protein–DNA complex in the asymmetric unit, with a solvent content of 62% and a Matthews coefficient of 3.22 Å{sup 3} Da{sup −1}.
- OSTI ID:
- 22360395
- Journal Information:
- Acta Crystallographica. Section F, Vol. 63, Issue Pt 8; Other Information: PMCID: PMC2335170; PMID: 17671361; PUBLISHER-ID: rp5005; OAI: oai:pubmedcentral.nih.gov:2335170; Copyright (c) International Union of Crystallography 2007; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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