Respiratory chain is required to maintain oxidized states of the DsbA-DsbB disulfide bond formation system in aerobically growing Escherichia coli cells.
DsbA, the disulfide bond catalyst of Escherichia coli, is a periplasmic protein having a thioredoxin-like Cys-30-Xaa-Xaa-Cys-33 motif. The Cys-30-Cys-33 disulfide is donated to a pair of cysteines on the target proteins. Although DsbA, having high oxidizing potential, is prone to reduction, it is maintained essentially all oxidized in vivo. DsbB, an integral membrane protein having two pairs of essential cysteines, reoxidizes DsbA that has been reduced upon functioning. It is not known, however, what might provide the overall oxidizing power to the DsbA-DsbB disulfide bond formation system. We now report that E. coli mutants defective in the hemA gene or in the ubiA-menA genes markedly accumulate the reduced form of DsbA during growth under the conditions of protoheme deprivation as well as ubiquinone/menaquinone deprivation. Disulfide bond formation of beta-lactamase was impaired under these conditions. Intracellular state of DsbB was found to be affected by deprivation of quinones, such that it accumulates first as a reduced form and then as a form of a disulfide-linked complex with DsbA. This is followed by reduction of the bulk of DsbA molecules. These results suggest that the respiratory electron transfer chain participates in the oxidation of DsbA, by acting primarily on DsbB. It is remarkable that a cellular catalyst of protein folding is connected to the respiratory chain.
Kobayashi
T
T
Institute for Virus Research, Kyoto University, Kyoto 606-01, Japan.
Kishigami
S
S
Sone
M
M
Inokuchi
H
H
Mogi
T
T
Ito
K
K
D000332:Aerobiosis; D000445:Aldehyde Oxidoreductases / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism*; D004579:Electron Transport; D004926:Escherichia coli / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D014451:Ubiquinone / Q000378:metabolism; D014812:Vitamin K / Q000378:metabolism
D001426:Bacterial Proteins; D004220:Disulfides; C079503:DsbB protein, Bacteria; D008565:Membrane Proteins; D014812:Vitamin K; D014451:Ubiquinone; D000445:Aldehyde Oxidoreductases; C067282:glutamyl tRNA reductase; D019704:Protein Disulfide-Isomerases