title: Protocols for Studying Protein Stability in an Arabidopsis Protoplast Transient Expression System.
abstract: Protein stability influences many aspects of biology, and measuring their stability in vivo can provide important insights into biological systems.This chapter describes in details two methods to assess the stability of a specific protein based on its transient expression in Arabidopsis protoplasts. First, a pulse-chase assay based on radioactive metabolic labeling of cellular proteins, followed by immunoprecipitation of the protein of interest. The decrease in radioactive signal is monitored over time and can be used to determine the protein's half-life.Alternatively, we also present a nonradioactive assay based on the use of reporter proteins, whose ratio can be quantified. This assay can be used to determine the relative stability of a protein of interest under specific conditions.
authors.affiliation: Laboratoire de Virologie Moléculaire, Institut Jacques Monod, CNRS, UMR 7592, Univ. Paris-Diderot, Sorbonne Paris Cité, Paris, France.|Adaptation des Plantes aux Contraintes Environnementales, UMR 7618, IEES Paris, Univ. Pierre-et-Marie-Curie, Paris, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Planchais
authors.affiliation: Laboratoire de Virologie Moléculaire, Institut Jacques Monod, CNRS, UMR 7592, Univ. Paris-Diderot, Sorbonne Paris Cité, Paris, France.|Laboratoire de Recherche en Sciences Végétales, CNRS, Univ. Toulouse Paul Sabatier, Toulouse, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Camborde
authors.affiliation: Laboratoire de Virologie Moléculaire, Institut Jacques Monod, CNRS, UMR 7592, Univ. Paris-Diderot, Sorbonne Paris Cité, Paris, France. isabelle.jupin@ijm.fr.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Jupin
mesh_terms: D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins / Q000737:chemistry*; D018506:Gene Expression Regulation, Plant; D047468:Immunoprecipitation; D008967:Molecular Biology / Q000379:methods*; D055550:Protein Stability; D011523:Protoplasts / Q000737:chemistry*; D014162:Transfection
chemical_list: D029681:Arabidopsis Proteins
keywords: Arabidopsis thaliana; Protein stability; Protoplasts; Pulse-chase; Transient expression; UPR assay