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
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Validation Study
MeSH terms
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14-3-3 Proteins / metabolism
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Cyclin-Dependent Kinase-Activating Kinase
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Cyclin-Dependent Kinases / metabolism
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Cyclins / metabolism
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DNA, Complementary / metabolism
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Mass Spectrometry
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Oryza / enzymology*
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Oryza / genetics
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Phosphorylation
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Plant Proteins / genetics
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Plant Proteins / isolation & purification
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Plant Proteins / metabolism*
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Plants, Genetically Modified / metabolism
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Protein Interaction Mapping / methods*
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Protein Kinases / genetics
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Protein Kinases / isolation & purification
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Protein Kinases / metabolism*
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Recombinant Fusion Proteins / isolation & purification
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Recombinant Fusion Proteins / metabolism
Substances
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14-3-3 Proteins
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Cyclins
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DNA, Complementary
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Plant Proteins
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Recombinant Fusion Proteins
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Protein Kinases
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Cyclin-Dependent Kinases
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Cyclin-Dependent Kinase-Activating Kinase