Figure 2.

Peroxidase activity of recombinant wild-type Prx2, with and without peroxynitrite treatment, and specific nitro variants. (A) Coupled assay of peroxidase activity of wild type Prx2. For activity measurements, 200 µM NADPH, 1 µM EcTR, 8 µM EcTrx1 and 0.5 µM Prx2 were used, and reaction was started with 10 µM H₂O₂ (black), 20 µM H₂O₂ (dark gray) or 50 µM H₂O₂ (light gray). Consumption of NADPH was followed at 340 nm. (B) Coupled assay peroxidase activity of nitrated wild-type Prx2 in the same conditions used in A. Inset. Coupled assay peroxidase activity of wild-type Prx2 non-treated (black) or treated with a 5-fold excess peroxynitrite (gray) using 10 µM H₂O₂. (C) Coupled assay peroxidase activity of nitro-variant NO₂Y33 Prx2 and (D) NO₂Y126 Prx2, in the same conditions used in A