Biochemical properties of the native PwGPx1 and PwGPx2 proteins. (A) The partially purified PwGPxs were resolved by 2-dimensional SDS-PAGE (15%, pH 3-10) and transferred onto nylon membranes. The membranes were reacted with the PwGPx-specific mouse antisera (1:2,000 dilutions). One-dimensional blots with PwGPxs (10 μg) and whole protein extracts (40 μg) of Paragonimus westermani (Pw-CE) and Clonorchis sinensis (Cs-CE) were also examined with a pooled mouse antiserum against C. sinensis PRx1 and PRx2 (1: 1,000 dilutions; our unpublished data). (B) The specificity and catalytic efficiency of the proteins purified by a series of gel chromatographies were examined toward catalytic substrates (H2O2 and cumene hydroperoxide [CHP]) and electron donors (glutathione [GSH] and thioredoxin [Trx]), respectively. The concentrations of GSH and Trx added in the reactions were empirically determined to ensure that the reductants are saturated. The absorbance values of experimental groups were substracted with those of reactions without PwGPxs. The reactions were performed in triplicate and the specific activities in μmol/min/mg were indicated as mean ± standard deviation.