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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Jarvis, P. (2008) Targeting of nucleus-encoded proteins to chloroplasts in plants. New Phytol. 179, 257–285.
Linka, N., and Weber, A. P. (2010) Intracellular metabolite transporters in plants. Mol. Plant 3, 21–53.
Block, M. A., Douce, R., Joyard, J., and Rolland, N. (2007) Chloroplast envelope membranes: a dynamic interface between plastids and the cytosol. Photosynth. Res. 92, 225–244
Joyard, J., Ferro, M., Masselon, C., Seigneurin-Berny, D., Salvi, D., Garin, J., and Rolland, N. (2009) Chloroplast proteomics and the compartmentation of plastidial isoprenoid biosynthetic pathways. Mol. Plant 2, 1154–1180.
Joyard, J., Ferro, M., Masselon, C., Seigneurin-Berny, D., Salvi, D., Garin, J., and Rolland, N. (2010) Chloroplast proteomics highlights the subcellular compartmentation of lipid metabolism. Prog. Lipid Res. 49, 128–158
Morsomme, P., Dambly, S., Maudoux, O., and Boutry, M. (1998) Single point mutations distributed in 10 soluble and membrane regions of the Nicotiana plumbaginifolia plasma membrane PMA2 H+−ATPase activate the enzyme and modify the structure of the C-terminal region. J. Biol. Chem. 273, 34837–34842.
Seigneurin-Berny, D., Gravot, A., Auroy, P., Mazard, C., Kraut, A., Finazzi, G., Grunwald, D., Rappaport, F., Vavasseur, A., Joyard, J., Richaud, P., and Rolland, N. (2006) HMA1, a new Cu-ATPase of the chloroplast envelope, is essential for growth under adverse light conditions. J. Biol. Chem. 281, 2882–2892.
Gerbeau, P., Guclu, J., Ripoche, P., and Maurel, C. (1999) Aquaporin Nt-TIPa can account for the high permeability of tobacco cell vacuolar membrane to small neutral solutes. Plant J. 18, 577–587.
Combettes, B., and Grienenberger, J. M. (1999) Analysis of wheat mitochondrial complex I purified by a one-step immunoaffinity chromatography. Biochimie 81, 645–653.
Perryman, R. A., Mooney, B., and Harmey, M. A. (1995) Identification of a 42-kDa plant mitochondrial outer membrane protein, MOM42, involved in the import of precursor proteins into plant mitochondria. Arch. Biochem. Biophys. 316, 659–664.
Vauclare, P., Macherel, D., Douce, R., and Bourguignon, J. (1998) The gene encoding T protein of the glycine decarboxylase complex involved in the mitochondrial step of the photorespiratory pathway in plants exhibits features of light induced genes. Plant Mol. Biol. 37, 309–318.
Miras, S., Salvi, D., Piette, L., Seigneurin-Berny, D., Grunwald, D., Garin, J., Reinbothe, C., Joyard, J., Reinbothe, S., and Rolland, N. (2007) Toc159- and Toc75-independent import of a transit sequence-less precursor into the inner envelope of chloroplasts. J. Biol. Chem. 282, 29482–29492.
Ferro, M., Brugière, S., Salvi, D., Seigneurin-Berny, D., Court, M., Moyet, L., Ramus, C., Miras, S., Mellal, M., Le Gall, S., Kieffer-Jaquinod, S., Bruley, C., Garin, J., Joyard, J., Masselon, C., and Rolland, N. (2010) AT_CHLORO, a comprehensive chloroplast proteome database with subplastidial localization and curated information on envelope proteins. Mol. Cell. Proteomics 9, 1063–1084.
Seigneurin-Berny, D., Salvi, D., Dorne, A.-J., Joyard, J., and Rolland, N. (2008) Percoll-purified and photosynthetically active chloroplasts from Arabidopsis thaliana leaves. Plant Physiol. Biochem. 46, 951–955.
Vallon, O., Bulte, L., Dainese P., Olive, J., Bassi, R., and Wollman, F. A. (1991) Lateral redistribution of cytochrome b6/f complexes along thylakoid membranes upon state transitions. Proc. Natl. Acad. Sci. USA 88, 8262–8266.
Ferro, M., Salvi, D., Brugière, S., Miras, S., Kowalski, S., Louwagie, M., Garin, J., Joyard, J., and Rolland, N. (2003) Proteomics of the chloroplast envelope membranes from Arabidopsis thaliana. Mol. Cell. Proteomics 2, 325–345.
Douce, R., and Joyard, J. (1982) Purification of the chloroplast envelope. In, Methods in Chloroplast Molecular Biology (Edelman, M., Hallick, R., and Chua, N.H., eds.) Elsevier, Amsterdam, North-Holland, pp. 139–256.
Carrie, C., Giraud, E., and Whelan, J. (2009) Protein transport in organelles: dual targeting of proteins to mitochondria and chloroplasts. FEBS J. 276, 1187–1195.
Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.
Seigneurin-Berny, D., Rolland, N., Garin, J., and Joyard, J. (1999) Differential extraction of hydrophobic proteins from chloroplast envelope membranes: a subcellular-specific proteomic approach to identify rare intrinsic membrane proteins. Plant J. 19, 217–228.
Ferro, M., Salvi, D., Rivière-Rolland, H., Vermat, T., Seigneurin-Berny, D., Grunwald, D., Garin J., Joyard, J., and Rolland, N. (2002) Integral membrane proteins of the chloroplast envelope: identification and subcellular localization of new transporters. Proc. Natl. Acad. Sci. USA 99, 11487–11492.
Chua, N. H. (1980) Electrophoretic analysis of chloroplast proteins. Methods Enzymol. 69, 434–436.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Salvi, D., Moyet, L., Seigneurin-Berny, D., Ferro, M., Joyard, J., Rolland, N. (2011). Preparation of Envelope Membrane Fractions from Arabidopsis Chloroplasts for Proteomic Analysis and Other Studies. In: Jarvis, R. (eds) Chloroplast Research in Arabidopsis. Methods in Molecular Biology, vol 775. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-237-3_10
Download citation
DOI: https://doi.org/10.1007/978-1-61779-237-3_10
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-236-6
Online ISBN: 978-1-61779-237-3
eBook Packages: Springer Protocols