title: Determination of Protein Carbonylation and Proteasome Activity in Seeds.
abstract: 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.
authors.affiliation: Institut de Biologie Paris-Seine, Unité Mixte de Recherche 7622, Sorbonne Universités, 75005, Paris, France.|Biologie du Développement, Institut de Biologie Paris-Seine, Unité Mixte de Recherche 7622, Centre National de la Recherche Scientifique, 75005, Paris, France.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Xia
authors.affiliation: Institut de Biologie Paris-Seine, Unité Mixte de Recherche 7622, Sorbonne Universités, 75005, Paris, France.|Biologie du Développement, Institut de Biologie Paris-Seine, Unité Mixte de Recherche 7622, Centre National de la Recherche Scientifique, 75005, Paris, France.
authors.forename: Hayat
authors.initials: H
authors.lastname: El-Maarouf-Bouteau
authors.affiliation: Institut de Biologie Paris-Seine, Unité Mixte de Recherche 7622, Sorbonne Universités, 75005, Paris, France.|Biologie du Développement, Institut de Biologie Paris-Seine, Unité Mixte de Recherche 7622, Centre National de la Recherche Scientifique, 75005, Paris, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Bailly
authors.affiliation: Institut de Biologie Paris-Seine, Unité Mixte de Recherche 7622, Sorbonne Universités, 75005, Paris, France. patrice.meimoun@upmc.fr.|Biologie du Développement, Institut de Biologie Paris-Seine, Unité Mixte de Recherche 7622, Centre National de la Recherche Scientifique, 75005, Paris, France. patrice.meimoun@upmc.fr.
authors.forename: Patrice
authors.initials: P
authors.lastname: Meimoun
mesh_terms: D008967:Molecular Biology / Q000379:methods*; D046912:Multiprotein Complexes / Q000302:isolation & purification*; D018384:Oxidative Stress / Q000235:genetics; D010944:Plants / Q000737:chemistry / Q000235:genetics; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D050050:Protein Carbonylation / Q000235:genetics*; D017382:Reactive Oxygen Species / Q000737:chemistry; D012639:Seeds / Q000737:chemistry; D015398:Signal Transduction
chemical_list: D046912:Multiprotein Complexes; D017382:Reactive Oxygen Species; D046988:Proteasome Endopeptidase Complex
keywords: Carbonylation; Degradation; Proteasome activity; Protein oxidation; Seeds