Expression of bovine adrenodoxin in E. coli and site-directed mutagenesis of /2 Fe-2S/ cluster ligands

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Item Type:	Article
Title:	Expression of bovine adrenodoxin in E. coli and site-directed mutagenesis of /2 Fe-2S/ cluster ligands
Creators Name:	Uhlmann, H., Beckert, V., Schwarz, D. and Bernhardt, R.
Abstract:	Expression systems for adrenodoxin into the periplasm and the cytoplasm of E. coli have been developed as a prerequisite for site-directed mutagenesis studies. In both systems the /2Fe-2S/ cluster of the protein was correctly assembled, the cytoplasmic one gives, however, a tenfold higher expression level. To determine which of the five cysteines at positions 46, 52, 55, 92, and 95 coordinate the /2Fe-2S/ center, they have been individually mutated into serines. From these mutants, only C95S forms a functionally active holoprotein. Thus, residues 46, 52, 55, and 92 are the cysteines that coordinate the /2Fe-2S/ cluster in adrenodoxin.
Keywords:	Adrenodoxin, Base Sequence, Biological Transport, Cysteine, Cytoplasm, DNA Mutational Analysis, Electron Spin Resonance Spectroscopy, Escherichia Coli, Gene Expression, Ligands, Molecular Cloning, Molecular Sequence Data, Recombinant Proteins, Serine, Site-Directed Mutagenesis, Structure-Activity Relationship, Transfection, Animals, Cattle
Source:	Biochemical and Biophysical Research Communications
ISSN:	0006-291X
Publisher:	Academic Press
Volume:	188
Number:	3
Page Range:	1131-1138
Date:	16 November 1992
Official Publication:	https://doi.org/10.1016/0006-291X(92)91349-U
PubMed:	View item in PubMed

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