Peculiar properties of DsbA in its export across the Escherichia coli cytoplasmic membrane.
Export of DsbA, a protein disulfide bond-introducing enzyme, across the Escherichia coli cytoplasmic membrane was studied with special reference to the effects of various mutations affecting translocation factors. It was noted that both the internalized precursor retaining the signal peptide and the periplasmic mature product fold rapidly into a protease-resistant structure and they exhibited anomalies in sodium dodecyl sulfate-polyacrylamide gel electrophoresis in that the former migrated faster than the latter. The precursor, once accumulated, was not exported posttranslationally. DsbA export depended on the SecY translocon, the SecA ATPase, and Ffh (signal recognition particle), but not on SecB. SecY mutations, such as secY39 and secY205, that severely impair translocation of a number of secretory substrates by interfering with SecA actions only insignificantly impaired the DsbA export. In contrast, secY125, affecting a periplasmic domain and impairing a late step of translocation, exerted strong export inhibition of both classes of proteins. These results suggest that DsbA uses not only the signal recognition particle targeting pathway but also a special route of translocation through the translocon, which is hence suggested to actively discriminate pre-proteins.
Shimohata
Nobuyuki
N
Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan.
Akiyama
Yoshinori
Y
Ito
Koreaki
K
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism* / Q000502:physiology; D008969:Molecular Sequence Data; D009154:Mutation; D014176:Protein Biosynthesis; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism; D021381:Protein Transport; D000069816:SEC Translocation Channels; D018271:Signal Recognition Particle / Q000502:physiology
D029968:Escherichia coli Proteins; D011498:Protein Precursors; D000069816:SEC Translocation Channels; C057841:SecY protein, E coli; D018271:Signal Recognition Particle; D019704:Protein Disulfide-Isomerases