KCS

Phosphorylation and dephosphorylation of protein kinases are important in signal transduction and cell cycle progression. Especially, some kinases like Cdc2 (same as Cdk-1) or MAPK (Mitogen-activated protein kinase) have double-phosphorylated states for its functional activation or inactivation. Cdc2 is an universal regulator protein of cell cycle progression pathway (G2 to M phase) in eukaryotic and prokaryotic cells and mitogen-activated protein (MAP) kinases are protein kinases that respond to extracellular stimuli (mitogens) and regulate various cellular activities, such as gene expression, mitosis, differentiation, and cell survival/apoptosis. So, scrutinizing of phosphorylated states of these kinases are crucial for accruate disease diagnosis related to these kinases. Herein, we show very selective fluorescent chemosensing of kinases using PyDPA. These chemosensor could clearly discriminate double phosphorylated states from mono-phosphorylated states of kinase. Using PyDPA, furthermore, we could make an successful application of real-time mornitoring of VHR phosphatase which efficiently dephosphorylate double-phosphorylated MAPK peptides.