title: A bioanalytical method for the proteome wide display and analysis of protein complexes from whole plant cell lysates.
abstract: While protein interaction studies and protein network modeling come to the forefront, the isolation and identification of protein complexes in a cellular context remains a major challenge for plant science. To this end, a nondenaturing extraction procedure was optimized for plant whole cell matrices and the combined use of gel filtration and BN-PAGE for the separation of protein complexes was studied. Hyphenation to denaturing electrophoresis and mass spectrometric analysis allows for the simultaneous identification of multiple (previously unidentified) protein interactions in single samples. The reliability and efficacy of the technique was confirmed (i) by the identification of well-studied plant protein complexes, (ii) by the presence of nonplant interologs for several of the novel complexes (iii) by presenting physical evidence of previously hypothetical plant protein interactions and (iv) by the confirmation of found interactions using co-IP. Furthermore practical issues concerning the use of this 2-D BN/SDS-PAGE display method for the analysis of protein-protein interactions are discussed.
authors.affiliation: Center for Proteome Analysis and Mass Spectrometry, University of Antwerp, Antwerp, Belgium.
authors.forename: Noor
authors.initials: N
authors.lastname: Remmerie
authors.forename: Luc
authors.initials: L
authors.lastname: Roef
authors.forename: Eveline
authors.initials: E
authors.lastname: Van De Slijke
authors.forename: Jelle
authors.initials: J
authors.lastname: Van Leene
authors.forename: Geert
authors.initials: G
authors.lastname: Persiau
authors.forename: Dominique
authors.initials: D
authors.lastname: Eeckhout
authors.forename: Hilde
authors.initials: H
authors.lastname: Stals
authors.forename: Kris
authors.initials: K
authors.lastname: Laukens
authors.forename: Filip
authors.initials: F
authors.lastname: Lemière
authors.forename: Eddy
authors.initials: E
authors.lastname: Esmans
authors.forename: Harry
authors.initials: H
authors.lastname: Van Onckelen
authors.forename: Dirk
authors.initials: D
authors.lastname: Inzé
authors.forename: Geert
authors.initials: G
authors.lastname: De Jaeger
authors.forename: Erwin
authors.initials: E
authors.lastname: Witters
mesh_terms: D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D047468:Immunoprecipitation; D010940:Plant Proteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis*
chemical_list: D010940:Plant Proteins; D020543:Proteome