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Methods to Analyze the Redox Reactivity of Plant Proteins

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Reactive Oxygen Species in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2526))

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Abstract

Proteins can be covalently modified by a broad range of highly reactive chemicals and redox mechanisms. Reversible redox-mediated post-translational modifications of sensitive cysteine thiol groups in proteins impact protein characteristics such as interaction behavior and activity state. Evaluating the response of proteins to redox perturbation or reactive chemical species is critical for understanding the underlying mechanisms involved and their contribution to plant stress physiology. Here we provide a detailed workflow that includes procedures for (i) purification, processing, and analysis of protein samples with redox agents, (ii) determining redox-modulated monomer to oligomer transitions using size exclusion chromatography, and (iii) activity assays for monitoring the impact of redox agents on purified enzymes and in crude extracts from plants subjected to oxidative stress. We exemplified how to apply several of the methods discussed for analyzing redox-sensing metallopeptidases, such as thimet oligopeptidases. We anticipate that these protocols should find broad applications in monitoring biochemical properties of other classes of redox-sensitive plant proteins.

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Acknowledgments

The authors acknowledge funding for this work from NSF-MCB 1714157 to SCP and GVP.

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Authors and Affiliations

  1. Department of Biochemistry, Molecular Biology, Entomology, and Plant Pathology, Mississippi State University, Mississippi State, MS, USA

    Thualfeqar Al-Mohanna, George V. Popescu & Sorina C. Popescu

  2. Institute for Genomics, Biocomputing, and Biotechnology, Mississippi State University, Mississippi State, MS, USA

    George V. Popescu

Authors
  1. Thualfeqar Al-Mohanna
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  2. George V. Popescu
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  3. Sorina C. Popescu
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Corresponding author

Correspondence to Sorina C. Popescu .

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Editors and Affiliations

  1. VIB Center for Plant Systems Biology, Ghent University, Zwijnaarde, Belgium

    Amna Mhamdi

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Al-Mohanna, T., Popescu, G.V., Popescu, S.C. (2022). Methods to Analyze the Redox Reactivity of Plant Proteins. In: Mhamdi, A. (eds) Reactive Oxygen Species in Plants. Methods in Molecular Biology, vol 2526. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2469-2_12

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  • DOI: https://doi.org/10.1007/978-1-0716-2469-2_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2468-5

  • Online ISBN: 978-1-0716-2469-2

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