Fig. 4: Differences in dimerization, thiol, and hydroperoxide consumption during the reaction of Prx2 with cis- versus trans-WOOH. | Nature Communications

Fig. 4: Differences in dimerization, thiol, and hydroperoxide consumption during the reaction of Prx2 with cis- versus trans-WOOH.

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

Fig. 4: Differences in dimerization, thiol, and hydroperoxide consumption during the reaction of Prx2 with cis- versus trans-WOOH.The alternative text for this image may have been generated using AI.

a Concentration-dependent reactions of cis- and trans-WOOH, and H2O2 (control) with pre-reduced His-tagged Prx2 (20 μM). Reactions were stopped after 10 s using NEM (50 mM). b Quantification of silver-stained Prx2 dimer and monomer bands in (a) with results expressed as ratio of Prx2 dimer-to-total Prx2, n = 4 independent experiments. c Quantification of free thiols after incubation of pre-reduced His-tagged Prx2 (20 μM) with cis-WOOH or trans-WOOH for 10 s before addition to 1 mM 5,5′-dithio-bis(2-nitrobenzoic acid), DTNB, n = 3 independent experiments. d Quantification of WOOH consumption and WOH formation in the presence of pre-reduced His-tagged Prx2 (20 μM) as determined by LC-MS/MS. Data are expressed as percentage of hydroperoxide consumed and corresponding alcohol formed relative to the initial hydroperoxide concentration, n = 3 independent experiments. e Absence of hyperoxidation of His-tagged Prx2 (20 μM) upon reaction with cis-WOOH or trans-WOOH. Samples were assessed in duplicates using non-reducing SDS-PAGE, with proteins on one gel silver stained and the other transferred to a polyvinylidene difluoride membrane before probing with an antibody specific for Prx-SO2/3. Data shown are representative of three (a) or two (e) separate experiments. Data in b, c, and d are shown as mean ± SEM. Uncropped blots and source data are provided as a Source data file.

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