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

Plant J. 2006 Apr;46(1):1-13. doi: 10.1111/j.1365-313X.2006.02671.x.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Validation Study

MeSH terms

  • 14-3-3 Proteins / metabolism
  • Cyclin-Dependent Kinases / metabolism
  • Cyclins / metabolism
  • DNA, Complementary / metabolism
  • Mass Spectrometry
  • Oryza / enzymology*
  • Oryza / genetics
  • Phosphorylation
  • Plant Proteins / genetics
  • Plant Proteins / isolation & purification
  • Plant Proteins / metabolism*
  • Plants, Genetically Modified / metabolism
  • Protein Interaction Mapping / methods*
  • Protein Kinases / genetics
  • Protein Kinases / isolation & purification
  • Protein Kinases / metabolism*
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism

Substances

  • 14-3-3 Proteins
  • Cyclins
  • DNA, Complementary
  • Plant Proteins
  • Recombinant Fusion Proteins
  • Protein Kinases
  • Cyclin-Dependent Kinases
  • cyclin-dependent kinase-activating kinase