Advanced search
1 file | 338.52 KB

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

Author
Organization
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.
Keywords
Arabidopsis, GFP, Immunoprecipitation, Protein complex identification, Protein–protein interaction, Tandem mass spectrometry

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 338.52 KB

Citation

Please use this url to cite or link to this publication:

Chicago
Wendrich, Jos, Sjef Boeren, Barbara Möller, Dolf Weijers, and Bert De Rybel. 2017. “In Vivo Identification of Plant Protein Complexes Using IP-MS/MS.” In Plant Hormones : Methods and Protocols, ed. Jürgen Kleine-Vehn and Michael Sauer, 1497:147–158. New York, NY, USA: Springer.
APA
Wendrich, J., Boeren, S., Möller, B., Weijers, D., & De Rybel, B. (2017). In Vivo Identification of Plant Protein Complexes Using IP-MS/MS. In J. Kleine-Vehn & M. Sauer (Eds.), Plant hormones : methods and protocols (Vol. 1497, pp. 147–158). New York, NY, USA: Springer.
Vancouver
1.
Wendrich J, Boeren S, Möller B, Weijers D, De Rybel B. In Vivo Identification of Plant Protein Complexes Using IP-MS/MS. In: Kleine-Vehn J, Sauer M, editors. Plant hormones : methods and protocols. New York, NY, USA: Springer; 2017. p. 147–58.
MLA
Wendrich, Jos, Sjef Boeren, Barbara Möller, et al. “In Vivo Identification of Plant Protein Complexes Using IP-MS/MS.” Plant Hormones : Methods and Protocols. Ed. Jürgen Kleine-Vehn & Michael Sauer. Vol. 1497. New York, NY, USA: Springer, 2017. 147–158. Print.
@incollection{8501699,
  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.},
  author       = {Wendrich, Jos and Boeren, Sjef and M{\"o}ller, Barbara and Weijers, Dolf and De Rybel, Bert},
  booktitle    = {Plant hormones : methods and protocols},
  editor       = {Kleine-Vehn, J{\"u}rgen and Sauer, Michael},
  isbn         = {9781493964673},
  issn         = {1064-3745},
  pages        = {147--158},
  publisher    = {Springer},
  series       = {Methods in Molecular Biology},
  title        = {In Vivo Identification of Plant Protein Complexes Using IP-MS/MS},
  url          = {http://dx.doi.org/10.1007/978-1-4939-6469-7\_14},
  volume       = {1497},
  year         = {2017},
}

Altmetric
View in Altmetric