作者
Anastassia Voronova, Jekaterina Kazantseva, Marina Tuuling, Niina Sokolova, Rannar Sillard, Peep Palumaa
发表日期
2007/5/1
期刊
Protein expression and purification
卷号
53
期号
1
页码范围
138-144
出版商
Academic Press
简介
Copper chaperone for cytochrome c oxidase (Cox17) is a 7kDa copper-binding protein, which facilitates incorporation of copper ions into CuA site of cytochrome c oxidase. Cox17 contains six conserved Cys residues and occurs in three different oxidative states, which display different metal-binding properties and stability. In the present study, we have elaborated technologies for production of partially oxidized human recombinant Cox17 in a bacterial expression system and purification of fully oxidized Cox17. For this purpose we used Escherichia coli Origami™ strain, which is deficient in thioredoxin and thioredoxin reductase systems and allows formation of disulfide bonds in cytoplasmic proteins. Fully oxidized Cox17 was purified by a simplified two-step procedure including gel filtration and cation exchange chromatography. By using mass spectrometry we demonstrated that application of 2-mercaptoethanol (2 …
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A Voronova, J Kazantseva, M Tuuling, N Sokolova… - Protein expression and purification, 2007