title: In Vivo Identification of Plant Protein Complexes Using IP-MS/MS.
abstract: Individual proteins often function as part of a protein complex. The identification of interacting proteins is therefore vital to understand the biological role and function of the studied protein. Here we describe a method for the in vivo identification of nuclear, cytoplasmic, and membrane-associated protein complexes from plant tissues using a strategy of immunoprecipitation followed by tandem mass spectrometry. By performing quantitative mass spectrometry measurements on biological triplicates, relative abundance of proteins in GFP-tagged complexes compared to background controls can be statistically evaluated to identify high-confidence interactors. We detail the entire workflow of this approach.
authors.affiliation: Laboratory of Biochemistry, Wageningen University, Dreijenlaan 3, 6703 HA, Wageningen, The Netherlands.
authors.forename: Jos R
authors.initials: JR
authors.lastname: Wendrich
authors.affiliation: Laboratory of Biochemistry, Wageningen University, Dreijenlaan 3, 6703 HA, Wageningen, The Netherlands.
authors.forename: Sjef
authors.initials: S
authors.lastname: Boeren
authors.affiliation: Laboratory of Biochemistry, Wageningen University, Dreijenlaan 3, 6703 HA, Wageningen, The Netherlands.|Department of Plant Systems Biology, VIB Ghent University, 9052, Ghent, Belgium.
authors.forename: Barbara K
authors.initials: BK
authors.lastname: Möller
authors.affiliation: Laboratory of Biochemistry, Wageningen University, Dreijenlaan 3, 6703 HA, Wageningen, The Netherlands. dolf.weijers@wur.nl.
authors.forename: Dolf
authors.initials: D
authors.lastname: Weijers
authors.affiliation: Laboratory of Biochemistry, Wageningen University, Dreijenlaan 3, 6703 HA, Wageningen, The Netherlands.|Department of Plant Systems Biology, Flemish Institute of Biotechnology, VIB Ghent University, 9052, Ghent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052, Ghent, Belgium.
authors.forename: Bert
authors.initials: B
authors.lastname: De Rybel
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D047468:Immunoprecipitation / Q000379:methods; D008565:Membrane Proteins / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D008565:Membrane Proteins; D010940:Plant Proteins
keywords: Arabidopsis; GFP; Immunoprecipitation; Protein complex identification; Protein–protein interaction; Tandem mass spectrometry