Fig. 3: Estimation of the rate constant for the reaction of cis- and trans-WOOH with His-tagged wild-type Prx2. | Nature Communications

Fig. 3: Estimation of the rate constant for the reaction of cis- and trans-WOOH with His-tagged wild-type Prx2.

From: Hydrogen peroxide signaling via its transformation to a stereospecific alkyl hydroperoxide that escapes reductive inactivation

Fig. 3: Estimation of the rate constant for the reaction of cis- and trans-WOOH with His-tagged wild-type Prx2.The alternative text for this image may have been generated using AI.

a Reaction of Prx2 (5 μM) with cis-WOOH (5–40 μM) at 25 °C. After incubation, N-ethylmaleimide (NEM, 30 mM) was added, and the samples subjected to non-reducing 4–12% SDS-PAGE followed by silver staining of the gels. b Time course of monomer disappearance with increasing concentrations of cis-WOOH. c Determination of second-order rate constant for the reaction or Prx2 with cis-WOOH (3.8 ± 0.5 × 103 M−1 s−1 at 25 °C). d Reaction of Prx2 (5 μM) with trans-WOOH (2–16 μM) at 5 °C. After incubation, NEM (30 mM) was added, and the samples subjected to non-reducing 4–12% SDS-PAGE followed by silver staining of the gels. Uncropped blots are provided in Source data. e Time course of Prx2 monomer disappearance with increasing concentrations of trans-WOOH. f Determination of the second-order rate constant for the reaction of Prx2 with trans-WOOH (1.1 ± 0.3 × 104 M−1 s−1 at 5 °C). Rate constants values are mean ± SEM. Results shown are representative of three independent experiments. Uncropped blots and source data are provided as a Source data file.

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