title: Protein complexes characterization in Arabidopsis thaliana by tandem affinity purification coupled to mass spectrometry analysis.
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.
authors.affiliation: URGV Plant Genomics, UMR INRA-1165/CNRS-ERL8196/Université d'Evry Val d'Essonne, 2, rue Gaston Crémieux, CP5708, 91057, Evry Cedex, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Bigeard
authors.forename: Delphine
authors.initials: D
authors.lastname: Pflieger
authors.forename: Jean
authors.initials: J
authors.lastname: Colcombet
authors.forename: Loïc
authors.initials: L
authors.lastname: Gérard
authors.forename: Hakim
authors.initials: H
authors.lastname: Mireau
authors.forename: Heribert
authors.initials: H
authors.lastname: Hirt
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D009532:Nickel / Q000737:chemistry; D059748:Proteolysis; D014357:Trypsin / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D009532:Nickel; D014357:Trypsin