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Peroxidatic cysteine residue of peroxiredoxin 2 separated from human red blood cells treated by tert-butyl hydroperoxide is hyperoxidized into sulfinic and sulfonic acids

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Abstract

Peroxiredoxin 2 (Prx2) is a redox enzyme that is abundantly expressed in red blood cells (RBCs) and has been the focus of clinical attention for monitoring the oxidative status. We previously developed a method to quantify the reduced and hyperoxidized forms of Prx2 in human RBCs using reverse-phase high-performance liquid chromatography (HPLC). In the present study, we investigated the hyperoxidative status of Prx2 at the molecular level in a post-translational modification analysis using a liquid chromatography–tandem mass spectrometry (LC–MS/MS) system. The LC–MS/MS analysis of the trypsin digests of Prx2 fractionated by reverse-phase HPLC demonstrated that the cysteine-51 residue (Cys-51) of the protein was modified with the hyperoxidative functional groups, sulfinic acid (–SO2H) and sulfonic acid (–SO3H), in RBCs treated with tert-butyl hydroperoxide (t-BHP). Furthermore, a selected ion monitoring (SIM) analysis quantitatively showed that sulfinic acid- and sulfonic acid-induced modifications in Prx2 Cys-51 were increased by the treatment with the oxidant. It was demonstrated that the peroxidatic cysteine of Prx2 separated using our HPLC system for oxidative monitoring was hyperoxidized into sulfinic acid and sulfonic acid in RBCs under an oxidative stress condition.

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Acknowledgements

We thank Mr. Shio Watanabe (Thermo Fisher Scientific Inc.) for their technical support.

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Authors and Affiliations

  1. Laboratory of Molecular and Cellular Biochemistry, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, 204-8588, Japan

    Yo-ichi Ishida & Masami Nagahama

  2. Department of Analytical Biochemistry, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, 204-8588, Japan

    Mariko Aki, Sohta Fujiwara & Yuki Ogasawara

Authors
  1. Yo-ichi Ishida
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  2. Mariko Aki
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  3. Sohta Fujiwara
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  4. Masami Nagahama
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  5. Yuki Ogasawara
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Correspondence to Yo-ichi Ishida or Yuki Ogasawara.

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Ishida, Yi., Aki, M., Fujiwara, S. et al. Peroxidatic cysteine residue of peroxiredoxin 2 separated from human red blood cells treated by tert-butyl hydroperoxide is hyperoxidized into sulfinic and sulfonic acids. Human Cell 30, 279–289 (2017). https://doi.org/10.1007/s13577-017-0171-0

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