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Isolation of transcription factor complexes from Arabidopsis cell suspension cultures by tandem affinity purification

Jelle Van Leene UGent, Dominique Eeckhout UGent, Geert Persiau UGent, Eveline Van De Slijke UGent, Jan Geerinck UGent, Gert Van Isterdael UGent, Erwin Witters and Geert De Jaeger UGent (2011) Plant transcription factors : methods and procotols. In Methods in Molecular Biology 754. p.195-218
abstract
Defining protein complexes is critical to virtually all aspects of cell biology because most cellular processes are regulated by stable or more dynamic protein interactions. Elucidation of the protein-protein interaction network around transcription factors is essential to fully understand their function and regulation. In the last decade, new technologies have emerged to study protein-protein interactions under near-physiological conditions. We have developed a high-throughput tandem affinity purification (TAP)/mass spectrometry (MS) platform for cell suspension cultures to analyze protein complexes in Arabidopsis thaliana. This streamlined platform follows an integrated approach comprising generic Gateway-based vectors with high cloning flexibility, the fast generation of transgenic suspension cultures, TAP adapted for plant cells, and tandem matrix-assisted laser desorption ionization MS for the identification of purified proteins. Recently, we evaluated the GS tag, originally developed to study mammalian protein complexes, that combines two IgG-binding domains of protein G with a streptavidin-binding peptide, separated by two tobacco etch virus cleavage sites. We found that this GS tag outperforms the traditional TAP tag in plant cells, regarding both specificity and complex yield. Here, we provide detailed protocols of the GS-based TAP platform that allowed us to characterize transcription factor complexes involved in signaling in response to the plant phytohormone jasmonate.
Please use this url to cite or link to this publication:
author
organization
year
type
bookChapter
publication status
published
subject
keyword
Arabidopsis thaliana, transcription factor, cell suspension culture, protein complex, tandem affinity purification, protein interaction
book title
Plant transcription factors : methods and procotols
editor
Ling Yuan and Sharyn E Perry
series title
Methods in Molecular Biology
volume
754
pages
195 - 218
publisher
Humana Press
place of publication
Totowa, NJ, USA
ISSN
1064-3745
ISBN
9781617791536
DOI
10.1007/978-1-61779-154-3_11
language
English
UGent publication?
yes
classification
B2
copyright statement
I have transferred the copyright for this publication to the publisher
id
1897817
handle
http://hdl.handle.net/1854/LU-1897817
date created
2011-09-06 12:44:02
date last changed
2011-09-07 10:51:47
@incollection{1897817,
  abstract     = {Defining protein complexes is critical to virtually all aspects of cell biology because most cellular processes are regulated by stable or more dynamic protein interactions. Elucidation of the protein-protein interaction network around transcription factors is essential to fully understand their function and regulation. In the last decade, new technologies have emerged to study protein-protein interactions under near-physiological conditions. We have developed a high-throughput tandem affinity purification (TAP)/mass spectrometry (MS) platform for cell suspension cultures to analyze protein complexes in Arabidopsis thaliana. This streamlined platform follows an integrated approach comprising generic Gateway-based vectors with high cloning flexibility, the fast generation of transgenic suspension cultures, TAP adapted for plant cells, and tandem matrix-assisted laser desorption ionization MS for the identification of purified proteins. Recently, we evaluated the GS tag, originally developed to study mammalian protein complexes, that combines two IgG-binding domains of protein G with a streptavidin-binding peptide, separated by two tobacco etch virus cleavage sites. We found that this GS tag outperforms the traditional TAP tag in plant cells, regarding both specificity and complex yield. Here, we provide detailed protocols of the GS-based TAP platform that allowed us to characterize transcription factor complexes involved in signaling in response to the plant phytohormone jasmonate.},
  author       = {Van Leene, Jelle and Eeckhout, Dominique and Persiau, Geert and Van De Slijke, Eveline and Geerinck, Jan and Van Isterdael, Gert and Witters, Erwin and De Jaeger, Geert},
  booktitle    = {Plant transcription factors : methods and procotols},
  editor       = {Yuan, Ling and Perry, Sharyn E},
  isbn         = {9781617791536},
  issn         = {1064-3745},
  keyword      = {Arabidopsis thaliana,transcription factor,cell suspension culture,protein complex,tandem affinity purification,protein interaction},
  language     = {eng},
  pages        = {195--218},
  publisher    = {Humana Press},
  series       = {Methods in Molecular Biology},
  title        = {Isolation of transcription factor complexes from Arabidopsis cell suspension cultures by tandem affinity purification},
  url          = {http://dx.doi.org/10.1007/978-1-61779-154-3\_11},
  volume       = {754},
  year         = {2011},
}

Chicago
Van Leene, Jelle, Dominique Eeckhout, Geert Persiau, Eveline Van De Slijke, Jan Geerinck, Gert Van Isterdael, Erwin Witters, and Geert De Jaeger. 2011. “Isolation of Transcription Factor Complexes from Arabidopsis Cell Suspension Cultures by Tandem Affinity Purification.” In Plant Transcription Factors : Methods and Procotols, ed. Ling Yuan and Sharyn E Perry, 754:195–218. Totowa, NJ, USA: Humana Press.
APA
Van Leene, J., Eeckhout, D., Persiau, G., Van De Slijke, E., Geerinck, J., Van Isterdael, G., Witters, E., et al. (2011). Isolation of transcription factor complexes from Arabidopsis cell suspension cultures by tandem affinity purification. In L. Yuan & S. E. Perry (Eds.), Plant transcription factors : methods and procotols (Vol. 754, pp. 195–218). Totowa, NJ, USA: Humana Press.
Vancouver
1.
Van Leene J, Eeckhout D, Persiau G, Van De Slijke E, Geerinck J, Van Isterdael G, et al. Isolation of transcription factor complexes from Arabidopsis cell suspension cultures by tandem affinity purification. In: Yuan L, Perry SE, editors. Plant transcription factors : methods and procotols. Totowa, NJ, USA: Humana Press; 2011. p. 195–218.
MLA
Van Leene, Jelle, Dominique Eeckhout, Geert Persiau, et al. “Isolation of Transcription Factor Complexes from Arabidopsis Cell Suspension Cultures by Tandem Affinity Purification.” Plant Transcription Factors : Methods and Procotols. Ed. Ling Yuan & Sharyn E Perry. Vol. 754. Totowa, NJ, USA: Humana Press, 2011. 195–218. Print.