Open AccessExpression, purification and X‐ray crystallographic analysis of thioredoxin from Streptomyces coelicolor
Author(s) -
Stefankova Petra,
Maderova Jana,
Barak Imrich,
Kollarova Marta,
Otwinowski Zbyszek
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309104032993
Subject(s) - streptomyces coelicolor , thioredoxin , escherichia coli , streptomyces , thioredoxin reductase , biology , biochemistry , bacteria , crystallography , chemistry , enzyme , gene , genetics
Thioredoxins are ubiquitous proteins that serve as reducing agents and general protein disulfide reductases. In turn, they are reduced by electrons obtained from the NADPH‐containing thioredoxin reductase. Thioredoxins have been isolated and characterized from a large number of organisms. The Gram‐positive bacterium Streptomyces coelicolor contains three thioredoxins that are involved in unknown biological processes. trxA from S. coelicolor was cloned and expressed in Escherichia coli and the protein purified and crystallized using the hanging‐drop method of vapour diffusion. The crystal structure of thioredoxin A has been determined at 1.5 Å resolution using a synchrotron‐radiation source. The protein reveals a thioredoxin‐like fold with a typical C XX C active site. The crystal exhibits the symmetry of space group P 2 1 2 1 2, with unit‐cell parameters a = 43.6, b = 71.8, c = 33.2 Å.