|Title||In vivo identification of plant protein complexes using IP-MS/MS|
|Author(s)||Wendrich, Jos R.; Boeren, Sjef; Möller, Barbara; Weijers, Dolf; Rybel, Bert de|
|Source||In: Methods in Molecular Biology / Kleine-Vehn, Jürgen, Sauer, Michael, Humana Press Inc. (Methods in Molecular Biology ) - ISBN 9781493964673 - p. 147 - 158.|
|Publication type||Peer reviewed book chapter|
|Keyword(s)||Arabidopsis - GFP - Immunoprecipitation - Protein complex identification - Protein–protein interaction - Tandem mass spectrometry|
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.