Abstract
Protein carbonylation is an irreversible oxidation process leading to a loss of function of carbonylated proteins. Carbonylation is largely considered as a hallmark of oxidative stress, the level of protein carbonylation being an indicator of the oxidative cellular status. The method described herein represents an adaptation to the commonly used 2,4-dinitrophenylhydrazine (DNPH)-based spectrophotometric method to monitor protein carbonylation level. The classical final sample precipitation was replaced by a gel filtration step avoiding the tedious and repetitive washings of the protein pellet to remove free DNPH while allowing optimal protein recovery.
This improved protocol here implemented to assay protein carbonylation in plant leaves can potentially be used with any cellular extract.
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Acknowledgments
This work was financially supported, thanks to the Poc in Labs initiative of the University Paris-Saclay and the Saclay Plant Sciences network, by the program “Investing for the Future” launched by the French State and implemented by the ANR (French National Research Agency). Former undergraduate students of the laboratory, Mai Pham Thy, Albert Kwarteng, and Oscar Freyschlag, are acknowledged for their participation to this work.
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Moreau, C., Issakidis-Bourguet, E. (2022). A Simplified Method to Assay Protein Carbonylation by Spectrophotometry. 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_10
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DOI: https://doi.org/10.1007/978-1-0716-2469-2_10
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