Chapter title |
In Vivo Identification of Plant Protein Complexes Using IP-MS/MS
|
---|---|
Chapter number | 14 |
Book title |
Plant Hormones
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6469-7_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6467-3, 978-1-4939-6469-7
|
Authors |
Wendrich, Jos R., Boeren, Sjef, Möller, Barbara K., Weijers, Dolf, De Rybel, Bert, Jos R. Wendrich, Sjef Boeren, Barbara K. Möller, Dolf Weijers, Bert De Rybel, Rybel, Bert |
Editors |
Jürgen Kleine-Vehn, Michael Sauer |
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. |
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