Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10537785 | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics | 2010 | 6 Pages |
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.
Keywords
Mitogen- and stress-activated protein kinase 1Bisubstrate inhibitorITCPKAcRIαGSTpKaH89EFCROCK-IITR-FRETHCsFITCYFPqPCRHTSPKCMSK1FRETChoFluorescence anisotropyBiosensorTamraChinese Hamster OvaryBinding assayNMRSurface plasmon resonanceSPRDissociation constantMass spectrometryNuclear magnetic resonance spectroscopyhigh-throughput screeningfluorescein isothiocyanatefluorescence polarizationArcinhibition constantHigh content screeningquantitative polymerase chain reactionFluorescent probeyellow fluorescent proteinProtein kinaseProtein kinase CcAMP-dependent protein kinaseIsothermal titration calorimetrycarboxytetramethylrhodamineglutathione S-transferase
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Asko Uri, Marje Lust, Angela Vaasa, Darja Lavogina, Kaido Viht, Erki Enkvist,