Chaperone activity of DsbC.
DsbC, a periplasmic disulfide isomerase of Gram-negative bacteria, displays about 30% of the activities of eukaryotic protein disulfide isomerase (PDI) as isomerase and as thiol-protein oxidoreductase. However, DsbC shows more pronounced chaperone activity than does PDI in promoting the in vitro reactivation and suppressing aggregation of denatured D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) during refolding. Carboxymethylation of DsbC at Cys98 decreases its intrinsic fluorescence, deprives of its enzyme activities, but lowers only partly its chaperone activity in assisting GAPDH reactivation. Simultaneous presence of DsbC and PDI in the refolding buffer shows an additive effect on the reactivation of GAPDH. The assisted reactivation of GAPDH and the protein disulfide oxidoreductase activity of DsbC can both be inhibited by scrambled and S-carboxymethylated RNases, but not by shorter peptides, including synthetic 10- and 14-mer peptides and S-carboxymethylated insulin A chain. In contrast, all the three peptides and the two nonnative RNases inhibit PDI-assisted GAPDH reactivation and the reductase activity of PDI. DsbC assists refolding of denatured and reduced lysozyme to a higher level than does PDI in phosphate buffer and does not show anti-chaperone activity in HEPES buffer. Like PDI, DsbC is also a disulfide isomerase with chaperone activity but may recognize different folding intermediates as does PDI.
National Laboratory of Biomacromolecules, Institute of Biophysics, Academia Sinica, Beijing 100101, China.
J
J
Chen
J L
JL
Song
S
S
Zhang
Y
Y
Wang
D F
DF
Cui
C C
CC
Wang
D001426:Bacterial Proteins / Q000378:metabolism; D004789:Enzyme Activation; D004926:Escherichia coli; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry; D019791:Guanidine / Q000494:pharmacology; D007700:Kinetics; D018832:Molecular Chaperones / Q000378:metabolism*; D009113:Muramidase / Q000737:chemistry; D010455:Peptides / Q000494:pharmacology; D011489:Protein Denaturation; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D017510:Protein Folding; D012260:Ribonucleases / Q000737:chemistry; D013050:Spectrometry, Fluorescence
D001426:Bacterial Proteins; D018832:Molecular Chaperones; D010455:Peptides; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D012260:Ribonucleases; D009113:Muramidase; D019704:Protein Disulfide-Isomerases; D019791:Guanidine