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

• Published in: The Plant journal : for cell and molecular biology 2006-04, Vol.46 (1), p.1-13
• Main Authors: Rohila, Jai S, Chen, Mei, Chen, Shuo, Chen, Johann, Cerny, Ronald, Dardick, Chris, Canlas, Patrick, Xu, Xia, Gribskov, Michael, Kanrar, Siddhartha, Zhu, Jian-Kang, Ronald, Pamela, Fromm, Michael E
• Format: Article
• Language: English
• Subjects: 14-3-3 Proteins - metabolism; affinity; Analytical, structural and metabolic biochemistry; Binding and carrier proteins; Biological and medical sciences; complementary DNA; Crop science; Cyclin-Dependent Kinases - metabolism; cyclins; Cyclins - metabolism; DNA, Complementary - metabolism; Enzymes; Fundamental and applied biological sciences. Psychology; Mass Spectrometry; Oryza - enzymology; Oryza - genetics; Oryza sativa; Phosphorylation; Plant Proteins - genetics; Plant Proteins - isolation & purification; Plant Proteins - metabolism; Plants, Genetically Modified - metabolism; progeny; Protein Interaction Mapping - methods; protein kinase; protein kinases; Protein Kinases - genetics; Protein Kinases - isolation & purification; Protein Kinases - metabolism; protein-protein interactions; Proteins; proteomics; Recombinant Fusion Proteins - isolation & purification; Recombinant Fusion Proteins - metabolism; Rice; tandem affinity purification; Transgenic plants
• ISSN: 0960-7412; 1365-313X
• DOI: 10.1111/j.1365-313X.2006.02671.x

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.