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

Authors

• Jai S. Rohila, University of Nebraska - Lincoln
• Mei Chen, University of Nebraska - LincolnFollow
• Shuo Chen, University of Nebraska - LincolnFollow
• Johann Chen, University of California, Davis
• Ronald CernyFollow
• Chris Dardick, University of California, Davis
• Patrick Canlas, University of California, DavisFollow
• Xia Xu, University of California, DavisFollow
• Michael Gribskov, Purdue UniversityFollow
• Siddhartha KanrarFollow
• Jian-Kang Zhu, University of California, Riverside
• P C. RonaldFollow
• Michael E. Fromm, University of Nebraska - LincolnFollow

Document Type

Article

Date of this Version

April 2006

Comments

Published in The Plant Journal 46:1 (2006), pp.1–13. doi: 10.1111/j.1365-313X.2006.02671.x Copyright © 2006 Jai S. Rohila, Mei Chen, Shuo Chen, Johann Chen, Ronald Cerny, Chris Dardick, Patrick Canlas, Xia Xu, Michael Gribskov, Siddhartha Kanrar, Jian-Kang Zhu, Pamela Ronald, and Michael E. Fromm. Published by Blackwell Publishing Ltd.

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.