title: Identification of a segment of DsbB essential for its respiration-coupled oxidation.
abstract: In the Escherichia coli protein disulphide bond formation pathway, membrane-bound DsbB oxidizes periplasmic DsbA, the disulphide bond-introducing enzyme. The Cys-41-Val-Leu-Cys-44 motif in the first periplasmic domain of DsbB is kept strongly oxidized by the respiratory function of the cell. We now show that the characteristic dithiothreitol resistance of the Cys-41-Cys-44 bond was retained even when the flanked Val-Leu combination was replaced by XX sequences from other oxidoreductases. Results of insertion mutagenesis showed that only the insertions (1-31 amino acids) in the region C-terminally adjacent to the CXXC motif impaired the oxidized state of DsbB. Deletion of a single amino acid from this region also rendered DsbB reduced and inactive. However, single amino acid substitutions of the four residues flanked by CXXC and the transmembrane segment did not abolish the oxidation of DsbB. These results suggest that some physical property, such as distance of the CXXC motif from the membrane, is important for the respiration-coupled oxidation of DsbB.
authors: Institute for Virus Research and CREST, Japan Science and Technology Corporation, Kyoto University, Kyoto 606-8507, Japan.
authors: T
authors: T
authors: Kobayashi
authors: Y
authors: Y
authors: Takahashi
authors: K
authors: K
authors: Ito
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D004352:Drug Resistance, Microbial; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D010101:Oxygen Consumption; D011487:Protein Conformation; D019704:Protein Disulfide-Isomerases / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C079503:DsbB protein, Bacteria; D008565:Membrane Proteins; D010088:Oxidoreductases; D019704:Protein Disulfide-Isomerases; D004229:Dithiothreitol