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Protein–protein interactions of tandem affinity purification‐tagged protein kinases in rice
Author(s) -
Rohila Jai S.,
Chen Mei,
Chen Shuo,
Chen Johann,
Cerny Ronald,
Dardick Chris,
Canlas Patrick,
Xu Xia,
Gribskov Michael,
Kanrar Siddhartha,
Zhu JianKang,
Ronald Pamela,
Fromm Michael E.
Publication year - 2006
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2006.02671.x
Subject(s) - tandem affinity purification , kinase , biology , genetically modified rice , phosphorylation , transgene , biochemistry , microbiology and biotechnology , gene , affinity chromatography , genetically modified crops , enzyme
Summary 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.