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Chapitre d'ouvrage

Separation, identification and profiling of membrane proteins by GFC/ IEC/SDS-PAGE and MALDI TOF MS

Abstract : Membrane protein identification by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) requires that proteins be separated prior to MS analysis. After membrane solubilization with the nondenaturing detergent n-dodecyl-beta-D-maltoside, proteins can be separated by ion-exchange chromatography (IEC) and further resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). An additional separation step by gel filtration (GF) before IEC/SDS-PAGE can be required depending on the complexity of the membrane protein mixture. Staining of final SDS-PAGE gels allows one to establish simply the protein expression pattern of a membrane fraction and to profile responses. Moreover, in-gel digestion of hydrophobic integral proteins is valuable. Finally, the resolution capacity of this separation procedure allows identification of proteins by MALDI-TOF MS. The method is illustrated by application to plant and yeast plasma membrane and to plant vacuolar membrane.
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Chapitre d'ouvrage
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https://hal.inrae.fr/hal-02821949
Déposant : Migration Prodinra <>
Soumis le : samedi 6 juin 2020 - 19:52:17
Dernière modification le : mercredi 1 juillet 2020 - 12:39:52

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  • HAL Id : hal-02821949, version 1
  • PRODINRA : 9739

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Wojciech Szponarski, Frederic Delom, Nicolas Sommerer, Michel Rossignol, Rémy Gibrat. Separation, identification and profiling of membrane proteins by GFC/ IEC/SDS-PAGE and MALDI TOF MS. Plant Proteomics : Methods and Protocols, 335, Humana Press, 2007, Methods in Molecular Biology, 978-1-59745-227-4. ⟨hal-02821949⟩

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