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Plant Cell Wall Proteomes: Bioinformatics and Cell Biology Tools to Assess the Bona Fide Cell Wall Localization of Proteins

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The Plant Cell Wall

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2149))

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Abstract

The purification of plant cell walls is challenging because they constitute an open compartment which is not limited by a membrane like the cell organelles. Different strategies have been established to limit the contamination by proteins of other compartments in cell wall proteomics studies. Non-destructive methods rely on washing intact cells with various types of solutions without disrupting the plasma membrane in order to elute cell wall proteins. In contrast, destructive protocols involve the purification of cell walls prior to the extraction of proteins with salt solutions. In both cases, proteins known to be intracellular have been identified by mass spectrometry in cell wall proteomes. The aim of this chapter is to provide tools to assess the subcellular localization of the proteins identified in cell wall proteomics studies, including: (1) bioinformatic predictions, (2) immunocytolocalization of proteins of interest on tissue sections and (3) in muro observation of proteins of interest fused to reporter fluorescent proteins by confocal microscopy. Finally, a qualitative assessment of the work can be performed and the strategy used to prepare the samples can be optimized if necessary.

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Acknowledgments

The authors are grateful to Université Paul Sabatier (Toulouse III), Université de Strasbourg and CNRS for support. They also wish to thank François Berthold and Caroline Hemmer for their great contribution to the cloning of pEAQ derivatives, Alain Jauneau for providing access to cell imaging facilities (http://trigenotoul.com/), and the bioinformatics platform of GenoToul Midi-Pyrénées for providing calculation facilities (http://bioinfo.genotoul.fr/).

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Authors and Affiliations

  1. Laboratoire de Recherche en Sciences Végétales, UMR 5546 Université Paul Sabatier-Toulouse III/CNRS, Pôle de Biotechnologie Végétale, 24 chemin de Borderouge, Auzeville Tolosane, France

    David Roujol, Laurent Hoffmann, Hélène San Clemente, Vincent Burlat & Elisabeth Jamet

  2. INRA, SVQV UMR_A 1131, Université de Strasbourg, Colmar, France

    Corinne Schmitt-Keichinger

  3. Institut de Biologie Moléculaire des Plantes (IBMP), UPR2357 CNRS, Université de Strasbourg, Strasbourg, France

    Christophe Ritzenthaler

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  1. David Roujol
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  2. Laurent Hoffmann
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  3. Hélène San Clemente
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  4. Corinne Schmitt-Keichinger
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  5. Christophe Ritzenthaler
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  6. Vincent Burlat
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  7. Elisabeth Jamet
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Correspondence to Elisabeth Jamet .

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Editors and Affiliations

  1. Botany and Plant Science School of Natural Sciences, Ryan Institute for Environmental Marine and Energy Research School of Natural Sciences, The National University of Ireland Galway, Galway, Ireland

    Zoë A. Popper

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Roujol, D. et al. (2020). Plant Cell Wall Proteomes: Bioinformatics and Cell Biology Tools to Assess the Bona Fide Cell Wall Localization of Proteins. In: Popper, Z. (eds) The Plant Cell Wall. Methods in Molecular Biology, vol 2149. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0621-6_25

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  • DOI: https://doi.org/10.1007/978-1-0716-0621-6_25

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0619-3

  • Online ISBN: 978-1-0716-0621-6

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