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Protein Complexes Characterization in Arabidopsis thaliana by Tandem Affinity Purification Coupled to Mass Spectrometry Analysis

  • Jean Bigeard
  • Delphine Pflieger
  • Jean Colcombet
  • Loïc Gérard
  • Hakim Mireau
  • Heribert HirtEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1171)

Abstract

Proteins are major elements participating in all the key functions of the cells. They rarely fulfill their physiological roles in an autonomous way but rather act as part of more complex cellular machines. Indeed they can bind different types of molecules (proteins, nucleic acids, metabolites, etc.), via stable or transient interactions, depending on their nature and functions. The identification of the molecular partners of a given protein is hence essential to better understand its roles, regulation, and mechanisms of action.

This chapter describes the use of a tandem affinity purification approach followed by mass spectrometry analysis to try to identify and characterize the proteins involved in protein complexes in Arabidopsis thaliana and decipher some mechanisms of regulation of the modules. Important elements to consider in such an approach are first extensively exposed in the introduction. This technique, in combination with complementary approaches like yeast two-hybrid and bimolecular fluorescence complementation, can be an interesting source of data to identify and characterize in vivo protein complexes.

Key words

Arabidopsis thaliana Protein complexes Tandem affinity purification Mass spectrometry Posttranslational modifications 

Notes

Acknowledgements

This work was partly funded by the French plant genomics program, Génoplante.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Jean Bigeard
    • 1
  • Delphine Pflieger
    • 2
  • Jean Colcombet
    • 1
  • Loïc Gérard
    • 1
  • Hakim Mireau
    • 3
  • Heribert Hirt
    • 1
    Email author
  1. 1.URGV Plant GenomicsUMR INRA-1165/CNRS-ERL8196/Université d’Evry Val d’EssonneEvry CedexFrance
  2. 2.Laboratoire Analyse et Modélisation pour la Biologie et l’EnvironnementCNRS UMR 8587, Université d’Evry Val d’EssonneEvry CedexFrance
  3. 3.Institut Jean-Pierre Bourgin, UMR1318 INRA-AgroParisTech, INRA Centre de Versailles-GrignonVersailles CedexFrance

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