Characterization of two alkyl hydroperoxide reductase C homologs alkyl hydroperoxide reductase C_H1 and alkyl hydroperoxide reductase C_H2 in Bacillus subtilis

Figure 4 Non-reducing SDS-PAGE of peroxiredoxins reduced with the Trx system. (A, B) AhpC_H1 and AhpC H2 were incubated without or with 2 mnol/L TCEP (A) or Trx system (B) for 30 min at 30 °C, followed by incubation with AMS for 30 min at 30 °C. The reaction was terminated by adding SDS sample buffer, and proteins were separated in 16% non-reducing SDS-PAGE gels. Protein migration pattern in A is the same as in B, suggesting that the Trx system reduced disulfide bonds in both proteins; d and m mark the position of dimeric and monomeric, respectively, AhpC_H1 and AhpC_H2. T1 and T2 denote the oxidized and reduced/modified Trx 1 proteins, respectively. Middle lanes show molecular weight markers (10, 15, 20, 25, 37, 50, 75, 100, 150, and 250 kDa). Trx, the Trx system; (C-F) Conversion of different Prxs from the oxidized to reduced form by the Trx system. 1, 2, 3, and 4 indicate molecular weight markers of 50, 37, 25, and 20 kDa, respectively.