Isolation of Plant Cell Wall Proteins

  • Elisabeth Jamet
  • Georges Boudart
  • Giséle Borderies
  • Stephane Charmont
  • Claude Lafitte
  • Michel Rossignol
  • Herve Canut
  • Rafael Pont-Lezica
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 425)

Summary

The quality of a proteomic analysis of a cell compartment strongly depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific drawbacks: (1) the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP) during the isolation procedure; (2) polysaccharide networks of cellulose, hemicelluloses, and pectins form potential traps for contaminants such as intracellular proteins; (3) the presence of proteins interacting in many different ways with the polysaccharide matrix require different procedures to elute them from the cell wall. Three categories of CWP are distinguished: labile proteins that have little or no interactions with cell wall components, weakly bound proteins extractable with salts, and strongly bound proteins. Two alternative protocols are decribed for cell wall proteomics: (1) nondestructive techniques allowing the extraction of labile or weakly bound CWP without damaging the plasma membrane; (2) destructive techniques to isolate cell walls from which weakly or strongly bound CWP can be extracted. These protocols give very low levels of contamination by intracellular proteins. Their application should lead to a realistic view of the cell wall proteome at least for labile and weakly bound CWP extractable by salts.

Key Words

Arabidopsis thaliana bioinformatics cell fractionation cell wall cell wall protein plant proteomics 
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Notes

Acknowledgments

The authors are grateful to the Universit’e Paul Sabatier (Toulouse III, France) and the CNRS for support.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Elisabeth Jamet
    • 1
  • Georges Boudart
    • 1
  • Giséle Borderies
    • 1
  • Stephane Charmont
    • 2
  • Claude Lafitte
    • 1
  • Michel Rossignol
    • 1
  • Herve Canut
    • 1
  • Rafael Pont-Lezica
    • 1
  1. 1.UMR 5546 CNRS-Université Paul Sabatier-Toulouse IIICastanet-TolosanFrance
  2. 2.Novartis Pharma AGBaselSwitzerland

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