Advanced search
1 file | 338.52 KB Add to list

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:

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.
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.
Chicago author-date
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.
Chicago author-date (all authors)
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.
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.
IEEE
[1]
J. Wendrich, S. Boeren, B. Möller, D. Weijers, and B. De Rybel, “In Vivo Identification of Plant Protein Complexes Using IP-MS/MS,” in Plant hormones : methods and protocols, vol. 1497, J. Kleine-Vehn and M. Sauer, Eds. New York, NY, USA: Springer, 2017, pp. 147–158.
@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 Möller, Barbara and Weijers, Dolf and De Rybel, Bert}},
  booktitle    = {{Plant hormones : methods and protocols}},
  editor       = {{Kleine-Vehn, Jürgen and Sauer, Michael}},
  isbn         = {{9781493964673}},
  issn         = {{1064-3745}},
  keywords     = {{Arabidopsis,GFP,Immunoprecipitation,Protein complex identification,Protein–protein interaction,Tandem mass spectrometry}},
  language     = {{und}},
  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