Contributions of substrate binding to the catalytic activity of DsbC.

DsbA and DsbC are involved in protein disulfide bond formation in the periplasm of Gram-negative bacteria. The two proteins are thought to fulfill different functions in vivo, DsbA as a catalyst of disulfide bond formation and DsbC as a catalyst of disulfide bond rearrangement. To explore the basis of this catalytic complementarity, the reaction mechanism of DsbC has been examined using unstructured model peptides that contain only one or two cysteine residues as substrates. The reactions between the various forms of the peptide and DsbC occur at rates up to 10(6)-fold faster than those that involve glutathione and DsbC, and they were constrained to occur at only one sulfur atom of disulfide bonds involving the peptide. Mixed disulfide complexes of DsbC and the peptide were 10(4)-fold more stable than the corresponding mixed disulfides with glutathione. These observations suggest that noncovalent binding interactions occur between the peptide and DsbC, which contribute to the very rapid kinetics of substrate utilization. The interactions between DsbC and the peptide appear to be more substantial than those between DsbA and the same peptide. The differences in the reaction of the peptide at the active sites of DsbA and DsbC provide insight into why DsbC is the better catalyst of disulfide bond rearrangement and how the active site chemistry of these structurally related proteins has been adapted to fulfill complementary functions.