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. 2014 Aug 13;289(39):27004–27018. doi: 10.1074/jbc.M114.564104

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FIGURE 4. — Oxidative activity of Prx4 toward PDI in the presence of BPA. A, oxidative activity of Prx4 (0.4 μm) toward PDI (10 μm) in the presence of various concentrations of BPA was assessed by measuring the decrease in A₃₄₀. B, Prx4-PDI-mediated oxidative folding of BPTI. HPLC profiles indicate the time course of oxidative folding of BPTI catalyzed by Prx4-PDI in the absence (left) and presence (right) of BPA (40 μm). Folding was initiated by the addition of Prx4 (0.1 μm) and PDI (1 μm) to reduced BPTI (30 μm) in a buffer containing H₂O₂ (200 μm). At the indicated time points, reactions were quenched with HCl and analyzed by HPLC. GSH/GSSG redox reagents were not included in this assay. C, quantitative analysis of each folding species based on the HPLC profiles shown in B. The occupancy of each species was plotted as a function of refolding time.