Overexpression, purification, crystallization and preliminary X‐ray diffraction analysis of the pMV158‐encoded plasmid transcriptional repressor protein CopG

FEBS Letters

Espinosa, Manuel²  González, Ana³  Solà, Marı́a¹  del Solar, Gloria²  Pérez-Luque, Rosa¹  Acebo, Paloma²  Coll, Miquel¹  Alda, M.Teresa²  Gomis-Rüth, F.Xavier¹ 

[1] Centre d'Investigació i Desenvolupament, C.S.I.C., Jordi Girona, 18-26, 08034 Barcelona, Spain;Centro de Investigaciones Biológicas, C.S.I.C., Velázquez, 144, 28006 Madrid, Spain;European Molecular Biology Organization, c/o DESY, Notkestraße 85, 22603 Hamburg, Germany

关键词: Plasmid;    Transcriptional repressor protein;    CopG;    Crystallization;    X-ray analysis;   

DOI  :  10.1016/S0014-5793(98)00219-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Plasmid pMV158 encodes a 45 amino acid transcriptional repressor, CopG, which is involved in copy number control. A new procedure for overproduction and purification of the protein has been developed. The CopG protein thus obtained retained its ability to specifically bind to DNA and to repress its own promoter. Purified CopG protein has been crystallized using the sitting-drop vapor diffusion method. The crystals, belonging to orthorhombic space group C222₁ (cell constants a=67.2 Å, b=102.5 Å, c=40.2 Å), were obtained from a solution containing methylpentanediol, benzamidine and sodium chloride, buffered to pH 6.7. Complete diffraction data up to 1.6 Å resolution have been collected. Considerations about the Matthews parameter account for the most likely presence of three molecules in the asymmetric unit (2.27 ų/Da).

【 授权许可】

Unknown