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Methods Mol Biol. 2011;775:189-206. doi: 10.1007/978-1-61779-237-3_10.

Preparation of envelope membrane fractions from Arabidopsis chloroplasts for proteomic analysis and other studies.

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1
Laboratoire de Physiologie Cellulaire Végétale, CNRS, CEA, INRA, Université de Grenoble, Grenoble, France.

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.

PMID:
21863444
DOI:
10.1007/978-1-61779-237-3_10
[Indexed for MEDLINE]

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