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Plant Hormones pp 147-158 | Cite as

In Vivo Identification of Plant Protein Complexes Using IP-MS/MS

  • Jos R. Wendrich
  • Sjef Boeren
  • Barbara K. Möller
  • Dolf WeijersEmail author
  • Bert De Rybel
Part of the Methods in Molecular Biology book series (MIMB, volume 1497)

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.

Key words

Protein–protein interaction Arabidopsis Immunoprecipitation Tandem mass spectrometry Protein complex identification GFP 

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Jos R. Wendrich
    • 1
  • Sjef Boeren
    • 1
  • Barbara K. Möller
    • 1
    • 4
  • Dolf Weijers
    • 1
    Email author
  • Bert De Rybel
    • 1
    • 2
    • 3
  1. 1.Laboratory of BiochemistryWageningen UniversityWageningenThe Netherlands
  2. 2.Department of Plant Systems Biology, Flemish Institute of BiotechnologyVIB Ghent UniversityGhentBelgium
  3. 3.Department of Plant Biotechnology and BioinformaticsGhent UniversityGhentBelgium
  4. 4.Department of Plant Systems BiologyVIB Ghent UniversityGhentBelgium

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