A mutation in either dsbA or dsbB, a gene encoding a component of a periplasmic disulfide bond-catalyzing system, is required for high-level expression of the Bacteroides fragilis metallo-beta-lactamase, CcrA, in Escherichia coli.
The metallo-beta-lactamase gene, ccrA, from Bacteroides fragilis is functionally expressed in Escherichia coli only in the presence of a genomic mutation in iarA or iarB (increased ampicillin resistance), identified in this study as dsbA or dsbB, respectively. DsbA and DsbB are components of a periplasmic protein disulfide bond-catalyzing system. Data indicated that DsbA interacted with CcrA, creating aberrant disulfide bond linkages that render CcrA proteolytically unstable. Mutations in dsbA or dsbB permissive for CcrA expression eliminated or greatly reduced DsbA activity, allowing CcrA to assume a disulfide bond-free and proteolytically stable conformation.
Alksne
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Department of Molecular Biology, American Cyanamid Company, Pearl River, New York 10965.
Keeney
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Rasmussen
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BA
D000668:Ampicillin Resistance / Q000235:genetics*; D001426:Bacterial Proteins; D001441:Bacteroides fragilis / Q000201:enzymology* / Q000235:genetics; D015153:Blotting, Western; D004220:Disulfides / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D009154:Mutation; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D001618:beta-Lactamases / Q000096:biosynthesis / Q000235:genetics*
D001426:Bacterial Proteins; D004220:Disulfides; D011994:Recombinant Proteins; D001618:beta-Lactamases; C063912:carbapenemase