Dsb-insensitive expression of CcrA, a metallo-beta-lactamase from Bacteroides fragilis, in Escherichia coli after amino acid substitution at two cysteine residues within CcrA.
It has previously been shown that functional expression of CcrA, a metallo-beta-lactamase from Bacteroides fragilis, in Escherichia coli requires a mutation in either dsbA or dsbB, components of a periplasmic disulfide bond-catalyzing system. Site-directed mutagenesis resulting in the substitution of various amino acids for two of the three cysteine residues within CcrA allowed the expression of CcrA in a dsb+ background. This finding supports the hypothesis that DsbA creates aberrant disulfide bonds involving the Cys residues of CcrA.
Elksne
L E
LE
Discovery Research, Wyeth-Ayerst Research, Pearl River, New York 10965,USA.
Rasmussen
B A
BA
D001426:Bacterial Proteins; D001441:Bacteroides fragilis / Q000201:enzymology*; D003545:Cysteine / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D007535:Isomerases / Q000378:metabolism*; D008667:Metalloproteins / Q000235:genetics / Q000378:metabolism*; D019704:Protein Disulfide-Isomerases; D011994:Recombinant Proteins / Q000378:metabolism; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D008667:Metalloproteins; D011994:Recombinant Proteins; D001618:beta-Lactamases; C063912:carbapenemase; D007535:Isomerases; D019704:Protein Disulfide-Isomerases; D003545:Cysteine