Responsive microsecond-lifetime photoluminescent probes for analysis of protein kinases and their inhibitors

•Responsive luminescent probes (ARC-Lum) for protein kinase (PK) research•Kinetic and thermodynamic characterization of PK inhibitors in homogeneous format•Characterization of binding properties of tight-binding inhibitors of PKs•Cell membrane permeable ARC-Lum probes for monitoring PK activity in live cells

Responsive ARC-Lum probes were used for measurement of the concentration of active protein kinases (PKs) and determination of affinity of inhibitors of PKs. ARC-Lum probes incorporate thiophene or a selenophene heterocycle and a fluorophore conjugated to the lysine residue in the peptide fragment. In the complex with a PK, ARC-Lum probes emit long-lifetime (microsecond-scale) luminescence at the emission wavelengths of the fluorescent label if the complex is illuminated at the excitation wavelength of the thiophene- or selenophene-containing phosphorescence donors. Bisubstrate ARC-Lum probes bind with sub-nanomolar affinity with several PKs of the AGC group. This article is part of a Special Issue entitled: Inhibitors of Protein Kinases (2012).

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