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

Figure 2 SDS-PAGE analyses of AhpC_H1 and AhpC_H2 proteins and their mutants carrying Cys→Ser substitutions. The wild-type (lane W) and mutated AhpC_H2 and AhpC_H1, and E. coli AhpC proteins were separated in non-reducing (A) and reducing (B) 12% SDS-PAGE gels; first lanes show molecular weight markers (15, 20, 25, 37, 50, 75, 100, 150, and 250 kDa). TCEP/AMS-treated (C) and non-treated (D) wild-type (lane W) and mutated AhpC_H1 proteins were separated in non-reducing 14% SDS-PAGE gels. AhpC_H1 and its C166S mutant reduced or not with the Trx system containing Trx 1, Trx reductase (TR), and NADPH were modified with a sulfhydryl group-specific reagent AMS and separated in non-reducing 14% SDS-PAGE gels (E). a marks the position of a single AhpC_H1 band (W, lane 1) and double bands (W, lane 4). b marks three positions of the AhpC_H1 band after the treatments indicated at the bottom. First lanes in B1, B2, and C show molecular weight markers (10, 15, 20, 25, 37, 50, 75, 100, 150, 250 kDa).