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Chapitre D'ouvrage Année : 2016

Determination of Protein Carbonylation and Proteasome Activity in Seeds

Qiong Xia
  • Fonction : Auteur
Hayat El-Maarouf-Bouteau
  • Fonction : Auteur
Patrice Meimoun

Résumé

Reactive oxygen species (ROS) have been shown to be toxic but also function as signaling molecules in a process called redox signaling. In seeds, ROS are produced at different developmental stages including dormancy release and germination. Main targets of oxidation events by ROS in cell are lipids, nucleic acids, and proteins. Protein oxidation has various effects on their function, stability, location, and degradation. Carbonylation represents an irreversible and unrepairable modification that can lead to protein degradation through the action of the 20S proteasome. Here, we present techniques which allow the quantification of protein carbonyls in complex protein samples after derivatization by 2,4-dinitrophenylhydrazine (DNPH) and the determination proteasome activity by an activity-based protein profiling (ABPP) using the probe MV151. These techniques, routinely easy to handle, allow the rapid assessment of protein carbonyls and proteasome activity in seeds in various physiological conditions where ROS may act as signaling or toxic elements.
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Dates et versions

hal-01560074 , version 1 (11-07-2017)

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Qiong Xia, Hayat El-Maarouf-Bouteau, Christophe Bailly, Patrice Meimoun. Determination of Protein Carbonylation and Proteasome Activity in Seeds. Lois, L. Maria and Matthiesen, Rune. Plant Proteostasis, 1450, Springer New York, pp.205-212, 2016, Methods in Molecular Biology, 978-1-4939-3757-8. ⟨10.1007/978-1-4939-3759-2_16⟩. ⟨hal-01560074⟩
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