title: Preparation of envelope membrane fractions from Arabidopsis chloroplasts for proteomic analysis and other studies.
abstract: Plastids are semiautonomous organelles restricted to plants and protists. These plastids are surrounded by a double membrane system, or envelope. These envelope membranes contain machineries to import nuclear-encoded proteins, and transporters for ions or metabolites, but are also essential for a range of plastid-specific metabolisms. Targeted semiquantitative proteomic investigations have revealed specific cross-contaminations by other cell or plastid compartments that may occur during chloroplast envelope purification. This article describes procedures developed to recover highly purified envelope fractions starting from Percoll-purified Arabidopsis chloroplasts, gives an overview of possible cross-contaminations, provides some tricks to limit these cross-contaminations, and lists immunological markers and methods that can be used to assess the purity of the envelope fractions.
authors.affiliation: Laboratoire de Physiologie Cellulaire Végétale, CNRS, CEA, INRA, Université de Grenoble, Grenoble, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Salvi
authors.forename: Lucas
authors.initials: L
authors.lastname: Moyet
authors.forename: Daphné
authors.initials: D
authors.lastname: Seigneurin-Berny
authors.forename: Myriam
authors.initials: M
authors.lastname: Ferro
authors.forename: Jacques
authors.initials: J
authors.lastname: Joyard
authors.forename: Norbert
authors.initials: N
authors.lastname: Rolland
mesh_terms: D017360:Arabidopsis / Q000166:cytology*; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002458:Cell Fractionation / Q000379:methods*; D005591:Chemical Fractionation; D002736:Chloroplasts / Q000737:chemistry* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007425:Intracellular Membranes / Q000737:chemistry* / Q000378:metabolism*; D013058:Mass Spectrometry; D018515:Plant Leaves / Q000166:cytology; D011205:Povidone / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D012822:Silicon Dioxide / Q000737:chemistry
chemical_list: D015415:Biomarkers; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone