Abstract
The optimal kinase activity of plant receptor-like kinases (RLKs) is often regulated by autophosphorylation of specific sites. Many of these phosphorylated residues then serve as recruiting sites for downstream interacting proteins. Therefore, identification of the phosphosites can be an important first step in delineating the signaling network. This chapter describes a protocol for identification of phosphorylated residues by mass spectrometry as well as a protocol to determine if an interacting partner can be phosphorylated in vitro.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
De Smet I, Voss U, Jurgens G, Beeckman T (2009) Receptor-like kinases shape the plant. Nat Cell Biol 11:1166–1173
Shiu SH, Bleecker AB (2001) Plant receptor-like kinase gene family: diversity, function, and signaling. Sci STKE 2001:re22
Oh MH, Wu X, Clouse SD, Huber SC (2011) Functional importance of BAK1 tyrosine phosphorylation in vivo. Plant Signal Behav 6:400–405
Meyer MR, Lichti CF, Townsend RR, Rao AG (2011) Identification of in vitro autophosphorylation sites and effects of phosphorylation on the Arabidopsis CRINKLY4 (ACR4) receptor-like kinase intracellular domain: insights into conformation, oligomerization, and activity. Biochemist 50:2170–2186
Liu GZ, Pi LY, Walker JC, Ronald PC, Song WY (2002) Biochemical characterization of the kinase domain of the rice disease resistance receptor-like kinase XA21. J Biol Chem 277:20264–20269
Johnson LN, Lewis RJ (2001) Structural basis for control by phosphorylation. Chem Rev 101:2209–2242
Burza AM, Pekala I, Sikora J, Siedlecki P, Malagocki P et al (2006) Nicotiana tabacum osmotic stress-activated kinase is regulated by phosphorylation on Ser-154 and Ser-158 in the kinase activation loop. J Biol Chem 281:34299–34311
Schulze WX, Deng L, Mann M (2005) Phosphotyrosine interactome of the ErbB-receptor kinase family. Mol Syst Biol 1:42–55
Meyer MR, Shah S, Zang J, Rohrs H, Rao AG (2015) Evidence for intermolecular interactions between the intracellular domains of the Arabidopsis receptor like kinase ACR4, its homologs and the WOX5 transcription factor. PLoS One 10(3):e0118861
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Meyer, M.R., Shah, S., Gururaj Rao, A. (2017). Investigation of Autophosphorylation Sites of Plant Receptor Kinases and Phosphorylation of Interacting Partners. In: Aalen, R. (eds) Plant Receptor Kinases. Methods in Molecular Biology, vol 1621. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7063-6_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7063-6_12
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7062-9
Online ISBN: 978-1-4939-7063-6
eBook Packages: Springer Protocols