Meyer Matthew R, Shah Shweta, Gururaj Rao A
C2N Diagnostics, 20 South Sarah Street, St. Louis, MO, 63108, USA.
Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA, 50011, USA.
Methods Mol Biol. 2017;1621:121-130. doi: 10.1007/978-1-4939-7063-6_12.
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.
植物类受体激酶（RLK）的最佳激酶活性通常受特定位点的自磷酸化调节。许多这些磷酸化残基随后作为下游相互作用蛋白的募集位点。因此，磷酸化位点的鉴定可能是描绘信号网络的重要第一步。本章介绍了一种通过质谱鉴定磷酸化残基的方法，以及一种确定相互作用伴侣是否能在体外被磷酸化的方法。
