Insights into Kinome Perturbation During NLRP3 Inflammasome Activation Using an Isotope-Coded ATP-affinity Probe and Targeted Mass Spectrometry

摘要

Here, we developed and used a MRM-based kinome profiling assay for approximately 300 human protein and lipid kinases by monitoring specifically targeted peptides located at the ATP-binding sites of the kinases. We demonstrated that ～ 250 protein kinases (50% of the entire human kinome and more than 80% of human kinome in a single cell line) could be routinely quantified without extensive separation using multi-dimensional chromatography. We also found that during inflammasome activation, the expression profiles of two kinases were changed. Currently, our on-going discoverybased analysis of kinases labeled using our ICAP reagents for different cell lines will further expand our kinome library.