Identification of protein kinase substrates by proteomic approaches

摘要

This review describes the current status of proteomic approaches to identify kinase substrates,nwhich may lead to valuable medical applications. It guides the reader towards various methodsnusing 2DE and liquid chromatography-tandem mass spectrometry. Dynamic changes ofnphosphorylation during extracellular stimuli can be quantitatively monitored by bothntechnologies. Among appropriate prefractionation procedures, the purification ofnphosphoproteins and phosphopeptides is an absolute step for success. The temporal changenand stoichiometry of phosphorylation are the important criteria to evaluate the physiologicalnmeaning of the reaction. Kinase substrates can also be identified by in vitro phosphorylationnsystems employing protein arrays, fractionated lysates, genetically engineered kinases andnphage libraries. The final section contains an expert opinion on the current strategies and thenissues we are going to challenge in the next 5 years.