Protein-protein interactions of tandem affinity purified protein kinases from rice.
Eighty-eight rice (Oryza sativa) cDNAs encoding rice leaf expressed protein kinases (PKs) were fused to a Tandem Affinity Purification tag (TAP-tag) and expressed in transgenic rice plants. The TAP-tagged PKs and interacting proteins were purified from the T1 progeny of the transgenic rice plants and identified by tandem mass spectrometry. Forty-five TAP-tagged PKs were recovered in this study and thirteen of these were found to interact with other rice proteins with a high probability score. In vivo phosphorylated sites were found for three of the PKs. A comparison of the TAP-tagged data from a combined analysis of 129 TAP-tagged rice protein kinases with a concurrent screen using yeast two hybrid methods identified an evolutionarily new rice protein that interacts with the well conserved cell division cycle 2 (CDC2) protein complex.
Rohila
Jai S
JS
Plant Science Initiative, University of Nebraska, Lincoln, NE, USA. jai.rohila@sdstate.edu
Chen
Mei
M
Chen
Shuo
S
Chen
Johann
J
Cerny
Ronald L
RL
Dardick
Christopher
C
Canlas
Patrick
P
Fujii
Hiroaki
H
Gribskov
Michael
M
Kanrar
Siddhartha
S
Knoflicek
Lucas
L
Stevenson
Becky
B
Xie
Mingtang
M
Xu
Xia
X
Zheng
Xianwu
X
Zhu
Jian-Kang
JK
Ronald
Pamela
P
Fromm
Michael E
ME
D000595:Amino Acid Sequence; D055162:Biocatalysis; D002846:Chromatography, Affinity; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D012275:Oryza / Q000201:enzymology* / Q000235:genetics; D010766:Phosphorylation; D011485:Protein Binding; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D020798:Two-Hybrid System Techniques
D018076:DNA, Complementary; D011494:Protein Kinases
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