title: Reduction of the periplasmic disulfide bond isomerase, DsbC, occurs by passage of electrons from cytoplasmic thioredoxin.
abstract: The Escherichia coli periplasmic protein DsbC is active both in vivo and in vitro as a protein disulfide isomerase. For DsbC to attack incorrectly formed disulfide bonds in substrate proteins, its two active-site cysteines should be in the reduced form. Here we present evidence that, in wild-type cells, these two cysteines are reduced. Further, we show that a pathway involving the cytoplasmic proteins thioredoxin reductase and thioredoxin and the cytoplasmic membrane protein DsbD is responsible for the reduction of these cysteines. Thus, reducing potential is passed from cytoplasmic electron donors through the cytoplasmic membrane to DsbC. This pathway does not appear to utilize the cytoplasmic glutathione-glutaredoxin pathway. The redox state of the active-site cysteines of DsbC correlates quite closely with its ability to assist in the folding of proteins with multiple disulfide bonds. Analysis of the activity of mutant forms of DsbC in which either or both of these cysteines have been altered further supports the role of DsbC as a disulfide bond isomerase.
authors.affiliation: Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: A
authors.initials: A
authors.lastname: Rietsch
authors.forename: P
authors.initials: P
authors.lastname: Bessette
authors.forename: G
authors.initials: G
authors.lastname: Georgiou
authors.forename: J
authors.initials: J
authors.lastname: Beckwith
mesh_terms: D007611:Aprotinin / Q000096:biosynthesis; D001665:Binding Sites; D002451:Cell Compartmentation; D003545:Cysteine / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004579:Electron Transport; D004926:Escherichia coli / Q000201:enzymology*; D005980:Glutathione Reductase / Q000235:genetics; D008958:Models, Molecular; D009154:Mutation; D019897:Periplasm / Q000201:enzymology; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D017510:Protein Folding; D013880:Thioredoxin-Disulfide Reductase / Q000378:metabolism; D013879:Thioredoxins / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis
chemical_list: D013879:Thioredoxins; D007611:Aprotinin; D005980:Glutathione Reductase; D013880:Thioredoxin-Disulfide Reductase; D014568:Urokinase-Type Plasminogen Activator; D019704:Protein Disulfide-Isomerases; D003545:Cysteine