Time-resolved luminescence detection of spleen tyrosine kinase activity through terbium sensitization.

ABSTRACT: Disruption of regulatory protein phosphorylation can lead to disease and is particularly prevalent in cancers. Inhibitors that target deregulated kinases are therefore a major focus of chemotherapeutic development. Achieving sensitivity and specificity in high-throughput compatible kinase assays is key to successful inhibitor development. Here, we describe the application of time-resolved luminescence detection to the direct sensing of spleen tyrosine kinase (Syk) activity and inhibition using a novel peptide substrate. Chelation and luminescence sensitization of Tb(3+) allowed the direct detection of peptide phosphorylation without any antibodies or other labeling reagents. Characterizing the Tb(3+) coordination properties of the phosphorylated vs unphosphorylated form of the peptide reve

SUBMITTER: Lipchik AM 

PROVIDER: S-EPMC3632334 | biostudies-literature | 2013 Mar

REPOSITORIES: biostudies-literature