title: In Vivo Radiolabeling of Arabidopsis Chloroplast Proteins and Separation of Thylakoid Membrane Complexes by Blue Native PAGE.
abstract: The investigation of membrane protein complex assembly and degradation is essential to understand cellular protein dynamics. Blue native PAGE provides a powerful tool to analyze the composition and formation of protein complexes. Combined with in vivo radiolabeling, the synthesis and decay of protein complexes can be monitored on a timescale ranging from minutes to several hours. Here, we describe a protocol to analyze thylakoid membrane complexes starting either with (35)S-methionine labeling of intact Arabidopsis leaves to investigate protein complex dynamics or with unlabeled leaf material to monitor steady-state complex composition.
authors.affiliation: Botanik, Department Biologie I, Ludwig-Maximilians-Universität, Großhaderner Strasse, 2-4, 82152, Planegg-Martinsried, Germany.
authors.forename: Catharina
authors.initials: C
authors.lastname: Nickel
authors.affiliation: Botanik, Department Biologie I, Ludwig-Maximilians-Universität, Großhaderner Strasse, 2-4, 82152, Planegg-Martinsried, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Brylok
authors.affiliation: Botanik, Department Biologie I, Ludwig-Maximilians-Universität, Großhaderner Strasse, 2-4, 82152, Planegg-Martinsried, Germany. serena.schwenkert@lmu.de.
authors.forename: Serena
authors.initials: S
authors.lastname: Schwenkert
mesh_terms: D017360:Arabidopsis / Q000737:chemistry; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D002736:Chloroplasts / Q000737:chemistry / Q000235:genetics; D046912:Multiprotein Complexes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D063347:Native Polyacrylamide Gel Electrophoresis / Q000379:methods*; D018515:Plant Leaves / Q000737:chemistry; D060431:Thylakoid Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D020524:Thylakoids / Q000737:chemistry / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; D046912:Multiprotein Complexes; D060431:Thylakoid Membrane Proteins
keywords: Arabidopsis thaliana; BN-PAGE; Photosynthetic protein complexes; Radiolabeling; Thylakoid membrane