Use of an oriented peptide library to determine the optimal substrates of protein kinases

摘要: Abstract Background: Phosphorylation by protein kinases is an important general mechanism for controlling intracellular processes, and plays essential part in the signal transduction pathways that regulate cell growth response to extracellular signals. A great number of have been discovered, identification their biological targets still a very active research area. Protein must appropriate substrate specificity ensure signals are transmitted correctly. Previous studies demonstrated importance primary sequences within proteins determining kinase specificity, but efficient ways identifying these lacking. Results We developed new technique kinases, using oriented library more than 2.5 billion peptide substrates. In this approach, consensus sequence optimal substrates determined sequencing mixture products generated during brief reaction with interest. The predicted cAMP-dependent (PKA) consistent known PKA cyclin-dependent (CDKs) cyclin B–Cdc2 A–CDK2 also agree well sites thought be phosphorylated vivo kinases. addition, we SLK1, homologue STE20 serine hitherto unknown specificity. discuss model incorporating into crystal structure kinase. Conclusion Using developed, can rapidly and, from information, potential identified.