Mf. Jeng et al., NMR CHARACTERIZATION OF A SINGLE-CYSTEINE MUTANT OF ESCHERICHIA-COLI THIOREDOXIN AND A COVALENT THIOREDOXIN-PEPTIDE COMPLEX, European journal of biochemistry, 257(2), 1998, pp. 299-308
The mechanism of disulfide reduction by thioredoxin in the cell is tho
ught to occur through the formation and subsequent destruction of a mi
xed-disulfide intermediate between thioredoxin and the substrate. In o
rder to model the interaction, we have prepared a mutant of Escherichi
a coli thioredoxin where the second cysteine residue of the active sit
e has been replaced by an alanine residue. A specific covalent complex
has been prepared between the remaining cysteine residue and a short
cysteine-containing peptide. This paper describes the preparation and
characterization of the mutant protein both free and in the peptide co
mplex.