Protein-protein interactions of tandem affinity purification-tagged protein kinases in rice.
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.
Plant Science Initiative, University of Nebraska, Lincoln, NE 68588, USA.
Jai S
JS
Rohila
Mei
M
Chen
Shuo
S
Chen
Johann
J
Chen
Ronald
R
Cerny
Chris
C
Dardick
Patrick
P
Canlas
Xia
X
Xu
Michael
M
Gribskov
Siddhartha
S
Kanrar
Jian-Kang
JK
Zhu
Pamela
P
Ronald
Michael E
ME
Fromm
D048948:14-3-3 Proteins / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D016213:Cyclins / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D013058:Mass Spectrometry; D012275:Oryza / Q000201:enzymology* / Q000235:genetics; D010766:Phosphorylation; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D011494:Protein Kinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D000095043:Cyclin-Dependent Kinase-Activating Kinase
D048948:14-3-3 Proteins; D016213:Cyclins; D018076:DNA, Complementary; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D011494:Protein Kinases; D018844:Cyclin-Dependent Kinases; D000095043:Cyclin-Dependent Kinase-Activating Kinase