Proteomics of loosely bound cell wall proteins of Arabidopsis thaliana cell suspension cultures: a critical analysis

Autor: Alain M. Boudet, Claude Lafitte, Bernard Monsarrat, Michel Rossignol, Gisèle Borderies, Rafael Pont-Lezica, Elisabeth Jamet, Alain Jauneau, Georges Boudart, Marie-Thérèse Esquerré-Tugayé
Přispěvatelé: Surfaces Cellulaires et Signalisation chez les Végétaux (SCSV), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Signalisation cellulaire et biotechnologie végétale (SCBV), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure Agronomique de Toulouse-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées, Institut de pharmacologie et de biologie structurale (IPBS), Génoplante (Contract N° NO 2001027), the Conseil National de la Recherche Scientifique and the Université Paul Sabatier, France, Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)-École nationale supérieure agronomique de Toulouse [ENSAT]-Université Toulouse III - Paul Sabatier (UT3), Institut National de la Recherche Agronomique (INRA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Supérieure Agronomique de Toulouse-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP)
Jazyk: angličtina
Rok vydání: 2003
Předmět:
0106 biological sciences
Arabidopsis thaliana
Clinical Biochemistry
cell suspension culture
Arabidopsis
MESH: Plant Extracts
MESH: Arabidopsis Proteins
Biology
Cell Fractionation
Proteomics
MESH: Research Support
Non-U.S. Gov't
01 natural sciences
Biochemistry
Analytical Chemistry
Cell wall
03 medical and health sciences
proteomics
MESH: Cell Wall
[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Genomics [q-bio.GN]
Electrophoresis
Gel
Two-Dimensional
MESH: Arabidopsis
Plant Proteins
030304 developmental biology
mass spectrometry
Gel electrophoresis
0303 health sciences
Chromatography
MESH: Plant Proteins
Arabidopsis Proteins
Plant Extracts
MESH: Proteomics
Extraction (chemistry)
Computational Biology
biology.organism_classification
MESH: Electrophoresis
Gel
Two-Dimensional
Apoplast
Membrane
Covalent bond
cell wall
MESH: Cell Fractionation
MESH: Spectrum Analysis
Mass
010606 plant biology & botany
MESH: Computational Biology
Zdroj: Electrophoresis
Electrophoresis, Wiley-VCH Verlag, 2003, 24 (19-20), pp.3421-32. ⟨10.1002/elps.200305608⟩
ISSN: 0173-0835
1522-2683
DOI: 10.1002/elps.200305608⟩
Popis: The complete sequencing of the Arabidopsis thaliana genome allows the use of the recently developed mass spectrometry techniques to identify the cell wall proteins (CWPs). Most proteomic approaches depend on the quality of sample preparation. Extraction of CWPs is particularly complex since the proteins may be free in the apoplast or are embedded in a polysaccharide matrix where they are retained by Van der Waals interactions, hydrogen bonds, hydrophobic or ionic interactions, or cross-linked by covalent bonds. Specific and sequential extraction procedures thus need to be developed. We report on the sequential extraction of loosely bound CWPs from living A. thaliana cells in culture. Different salts and chelating agents were used for releasing the proteins from the wall. Their effects on the extraction of CWPs and on the integrity of the plasma membrane were evaluated. Bioinformatic software was used to identify proteins and to predict their sub-cellular localization. The obtained data show that the plasma membrane of cells in culture was easily damaged by some steps of the extraction procedure, leading to the release of increasing amounts of intracellular proteins. Nevertheless, we identified fifty CWPs among which thirteen were new proteins for the cell wall. In addition, 76% of these CWPs were basic proteins not resolved in two-dimensional (2-D) gel electrophoresis. The existence of two hypothetical proteins was confirmed. The structure of three proteins could be confirmed using mass spectrometry data.
Databáze: OpenAIRE
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