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Monitoring of Methionine Sulfoxide Content and Methionine Sulfoxide Reductase Activity

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Selenoproteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1661))

Abstract

The sulfur-containing amino acid methionine (Met) plays critical roles in protein synthesis, methylation, and sulfur metabolism. Both in its free form and in the form of an amino acid residue, it can be oxidized to the R and S diastereomers of methionine sulfoxide (MetO). Organisms evolved methionine sulfoxide reductases (MSRs) to reduce MetO to Met, with the MSRs type A (MSRA) and type B (MSRB) being specific for the S and R forms of MetO, respectively. In mammals, the selenoprotein MSRB1 plays an important protein repair function, and its expression is tightly regulated by dietary selenium. In this chapter, we describe a protocol for determining the concentration of protein-based Met-R-O and its analysis in HEK293 cells using a genetically encoded ratiometric fluorescent biosensor MetROx. We also describe the procedure for quantifying MSR activities in cell extracts using specific substrates and a reverse phase HPLC-based method.

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Acknowledgments

This work was supported by the NIH grant AG021518. Pascal Rey (CEA, DRF, BIAM, Laboratoire d’Ecophysiologie Moléculaire des Plantes, France) is acknowledged for technical advice.

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Authors and Affiliations

  1. Laboratoire de Bioénergétique Cellulaire, Institut de Biosciences et Biotechnologies Aix-Marseille (BIAM), Commissariat à l‘Energie Atomique et aux Energies Alternatives (CEA), 13108, Saint-Paul-lès-Durance, France

    Lionel Tarrago

  2. UMR 7265, Centre National de Recherche Scientifique, Saint-Paul-lès-Durance, France

    Lionel Tarrago

  3. Aix Marseille Université, Marseille, France

    Lionel Tarrago

  4. Centrale Marseille, CNRS, iSm2 UMR 7313, Aix-Marseille Université, 13397, Marseille, France

    Emmanuel Oheix

  5. Division of Genetics, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA, 02115, USA

    Zalán Péterfi & Vadim N. Gladyshev

Authors
  1. Lionel Tarrago
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  2. Emmanuel Oheix
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  3. Zalán Péterfi
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  4. Vadim N. Gladyshev
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Corresponding authors

Correspondence to Lionel Tarrago or Vadim N. Gladyshev .

Editor information

Editors and Affiliations

  1. Centre International de Recherche en Infectiologie, CIRI, Lyon, France

    Laurent Chavatte

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Tarrago, L., Oheix, E., Péterfi, Z., Gladyshev, V.N. (2018). Monitoring of Methionine Sulfoxide Content and Methionine Sulfoxide Reductase Activity. In: Chavatte, L. (eds) Selenoproteins. Methods in Molecular Biology, vol 1661. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7258-6_20

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  • DOI: https://doi.org/10.1007/978-1-4939-7258-6_20

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7257-9

  • Online ISBN: 978-1-4939-7258-6

  • eBook Packages: Springer Protocols

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