Back to Search
Start Over
Purification of ArcR, an oxidation-sensitive regulatory protein from Bacillus licheniformis.
- Source :
-
Protein expression and purification [Protein Expr Purif] 2004 Sep; Vol. 37 (1), pp. 32-8. - Publication Year :
- 2004
-
Abstract
- In Bacillus licheniformis, ArcR, a transcriptional activator of the Crp/Fnr family, is required for expression of the anaerobic pathway of arginine catabolism, the arginine deiminase pathway. The method described here allows the purification of milligram quantities of functional ArcR from a recombinant Escherichia coli strain. The solubility properties of ArcR were much exploited during the purification process. The protein appeared highly sensitive to oxidation. Oxidation-induced precipitation of the protein was attributed to the formation of intermolecular disulfide bridges. Alkylation of mutant proteins with single substitutions showed that both cysteine residues of the protein, C178 and C205, are involved in formation of the disulfide bridges. Substitution of both cysteines yielded a functional protein insensitive to oxidation and able to form a complex with its cognate target on the DNA.
- Subjects :
- Amino Acid Sequence
Bacillus genetics
Bacillus metabolism
Bacterial Proteins genetics
Bacterial Proteins metabolism
Cloning, Molecular
Cysteine chemistry
DNA-Binding Proteins genetics
DNA-Binding Proteins metabolism
Disulfides chemistry
Molecular Sequence Data
Oxidation-Reduction
Recombinant Proteins genetics
Recombinant Proteins isolation & purification
Recombinant Proteins metabolism
Salts chemistry
Sequence Alignment
Bacillus chemistry
Bacterial Proteins isolation & purification
DNA-Binding Proteins isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 1046-5928
- Volume :
- 37
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Protein expression and purification
- Publication Type :
- Academic Journal
- Accession number :
- 15294278
- Full Text :
- https://doi.org/10.1016/j.pep.2004.05.006