title: Bisubstrate fluorescent probes and biosensors in binding assays for HTS of protein kinase inhibitors.
abstract: Conjugates of adenosine mimics and d-arginine-rich peptides (ARCs) are potent inhibitors of protein kinases (PKs) from the AGC group. Labeling ARCs with fluorescent dyes or immobilizing on chip surfaces gives fluorescent probes (ARC-Photo) and biosensors that can be used for high-throughput screening (HTS) of inhibitors of protein kinases. The bisubstrate character (simultaneous association with both binding sites of the kinase) and high affinity of ARCs allow ARC-based probes and sensors to be used for characterization of inhibitors targeted to either binding site of the kinase with affinities in whole nanomolar to micromolar range. The ability to penetrate cell plasma membrane and bind to the target kinase fused with a fluorescent protein leads to the possibility to use ARC-Photo probes for high content screening (HCS) of inhibitors in cellular milieu with detection of intensity of Förster resonance energy transfer (FRET) between two fluorophores.
authors.affiliation: Institute of Chemistry, University of Tartu, Ravila 14a, 50411 Tartu, Estonia. asko.uri@ut.ee
authors.forename: Asko
authors.initials: A
authors.lastname: Uri
authors.forename: Marje
authors.initials: M
authors.lastname: Lust
authors.forename: Angela
authors.initials: A
authors.lastname: Vaasa
authors.forename: Darja
authors.initials: D
authors.lastname: Lavogina
authors.forename: Kaido
authors.initials: K
authors.lastname: Viht
authors.forename: Erki
authors.initials: E
authors.lastname: Enkvist
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000378:metabolism; D011494:Protein Kinases / Q000737:chemistry*
chemical_list: D005456:Fluorescent Dyes; D010455:Peptides; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases