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Plant Proteomics pp 93–109Cite as

Plant Plasma Membrane Protein Extraction and Solubilization for Proteomic Analysis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 355))

Abstract

The plasma membrane (PM) exists as the interface between the cytosol and the environment in all living cells and is one of the most complex and differentiated membrane. The identification and characterization of membrane proteins (either extrinsic or intrinsic) is a crucial challenge since many of these proteins are involved in essential cellular functions such as cell signaling, osmoregulation, nutrition, and metabolism. Methods to isolate PM fractions vary according to organisms, tissues, and cell type. This chapter emphasizes isolation, from the model plant Arabidopsis thaliana, of PM fractions from a microsomal membrane fraction by two-phase partioning, a methodology that utilizes the different surface properties of membranes. PM proteins that do not span the lipid bilayer are generally well recovered after 2D gel electrophoresis. By contrast, the recovery of transmembrane proteins requires first the depletion of the PM fraction from soluble proteins, being either cytosolic contaminants or functionally associated proteins, and second, to the use of specific solubilization procedures. This chapter presents protocols to strip PM based on alkaline treatment of membranes and to solubilize hydrophobic proteins to increase their recovery on 2D gels. Aquaporins that are highly hydrophobic proteins are used to probe the relevance of the procedures.

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Author information

Authors and Affiliations

  1. Biochimie et Physiologie Moléculaire des Plantes, INRA, Montpellier, France

    Véronique Santoni

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  1. Véronique Santoni
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Editor information

Editors and Affiliations

  1. UMR de Génétique Végétale, La Ferme du Moulon, Gif-sur-Yvette, France

    Hervé Thiellement , Michel Zivy  & Catherine Damerval ,  & 

  2. UMR de Chimie Biologique, Thiverval Grignon, France

    Valérie Méchin

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© 2007 Humana Press Inc., Totowa, NJ

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Santoni, V. (2007). Plant Plasma Membrane Protein Extraction and Solubilization for Proteomic Analysis. In: Thiellement, H., Zivy, M., Damerval, C., Méchin, V. (eds) Plant Proteomics. Methods in Molecular Biology, vol 355. Humana Press. https://doi.org/10.1385/1-59745-227-0:93

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  • DOI: https://doi.org/10.1385/1-59745-227-0:93

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-635-1

  • Online ISBN: 978-1-59745-227-4

  • eBook Packages: Springer Protocols

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