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  • Title: Preparation of envelope membrane fractions from Arabidopsis chloroplasts for proteomic analysis and other studies.
    Author: Salvi D, Moyet L, Seigneurin-Berny D, Ferro M, Joyard J, Rolland N.
    Journal: Methods Mol Biol; 2011; 775():189-206. PubMed ID: 21863444.
    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.
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