title: Global kinase screening. Applications of frontal affinity chromatography coupled to mass spectrometry in drug discovery.
abstract: Utilizing frontal affinity chromatography with mass spectrometry detection (FAC-MS), we have identified novel applications in the discovery of small-molecule hits to protein targets that are difficult if not impossible to accomplish using traditional assays. We demonstrate for the first time an ability to distinguish between competitive ligands for the ATP and substrate sites of protein kinase C independently in the same experiment and show that ATP competitive ligands using a functionally inactive receptor tyrosine kinase can be identified. This ability of FAC-MS to simultaneously monitor binding at the ATP and substrate binding sites, as well as measure ligand binding to both active and inactive kinases, suggests that FAC-MS can be used as a "global kinase binding assay".
authors.affiliation: Lead Discovery and Optimization Division, Protana Inc., 251 Attwell Drive, Toronto, Ontario, M9W 7H4 Canada.
authors.forename: Jacek J
authors.initials: JJ
authors.lastname: Slon-Usakiewicz
authors.forename: Jin-Rui
authors.initials: JR
authors.lastname: Dai
authors.forename: William
authors.initials: W
authors.lastname: Ng
authors.forename: J Estelle
authors.initials: JE
authors.lastname: Foster
authors.forename: Eugen
authors.initials: E
authors.lastname: Deretey
authors.forename: Leticia
authors.initials: L
authors.lastname: Toledo-Sherman
authors.forename: Peter R
authors.initials: PR
authors.lastname: Redden
authors.forename: Andrew
authors.initials: A
authors.lastname: Pasternak
authors.forename: Neil
authors.initials: N
authors.lastname: Reid
mesh_terms: D000470:Alkaloids / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D053119:Benzophenanthridines / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D020134:Catalytic Domain; D002846:Chromatography, Affinity / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D007093:Imidazoles / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D015394:Molecular Structure; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D051571:Protein Kinase C-alpha / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D011494:Protein Kinases / Q000737:chemistry* / Q000378:metabolism; D011725:Pyridines / Q000737:chemistry / Q000378:metabolism; D011799:Quinazolines / Q000737:chemistry / Q000378:metabolism; D036183:Receptor, EphB2 / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D000470:Alkaloids; D053119:Benzophenanthridines; D007093:Imidazoles; D010455:Peptides; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D011799:Quinazolines; D011994:Recombinant Proteins; C118361:WHI P180; C016299:chelerythrine; D011494:Protein Kinases; D036183:Receptor, EphB2; D051571:Protein Kinase C-alpha; C093642:SB 203580; C088860:4-((3-bromophenyl)amino)-6,7-dimethoxyquinazoline