Protein-protein interactions of tandem affinity purification-tagged protein kinases in rice

Jai S. Rohila, Mei Chen, Shuo Chen, Johann Chen, Ronald Cerny, Chris Dardick, Patrick Canlas, Xia Xu, Michael Gribskov, Siddhartha Kanrar, Jian Kang Zhu, Pamela Ronald, Michael E. Fromm

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109 Scopus citations

Abstract

Forty-one rice cDNAs encoding protein kinases were fused to the tandem affinity purification (TAP) tag and expressed in transgenic rice plants. The TAP-tagged kinases and interacting proteins were purified from the T1 progeny of the transgenic rice plants and identified by mass spectrometry. Ninety-five percent of the TAP-tagged kinases were recovered. Fifty-six percent of the TAP-tagged kinases were found to interact with other rice proteins. A number of these interactions were consistent with known protein complexes found in other species, validating the TAP-tag method in rice plants. Phosphorylation sites were identified on four of the kinases that interacted with either 14-3-3 proteins or cyclins.

Original languageEnglish (US)
Pages (from-to)1-13
Number of pages13
JournalPlant Journal
Volume46
Issue number1
DOIs
StatePublished - Apr 1 2006

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Keywords

  • Affinity
  • Oryza sativa
  • Protein kinase
  • Proteomics
  • Tandem affinity purification

ASJC Scopus subject areas

  • Genetics
  • Plant Science
  • Cell Biology

Cite this

Rohila, J. S., Chen, M., Chen, S., Chen, J., Cerny, R., Dardick, C., Canlas, P., Xu, X., Gribskov, M., Kanrar, S., Zhu, J. K., Ronald, P., & Fromm, M. E. (2006). Protein-protein interactions of tandem affinity purification-tagged protein kinases in rice. Plant Journal, 46(1), 1-13. https://doi.org/10.1111/j.1365-313X.2006.02671.x