title: A three-hybrid approach to scanning the proteome for targets of small molecule kinase inhibitors.
abstract: In this study, we explored the application of a yeast three-hybrid (Y3H)-based compound/protein display system to scanning the proteome for targets of kinase inhibitors. Various known cyclin-dependent kinase (CDK) inhibitors, including purine and indenopyrazole analogs, were displayed in the form of methotrexate-based hybrid ligands and deployed in cDNA library or yeast cell array-based screening formats. For all inhibitors, known cell cycle CDKs as well as novel candidate CDK-like and/or CDK-unrelated kinase targets could be identified, many of which were independently confirmed using secondary enzyme assays and affinity chromatography. The Y3H system described here may prove generally useful in the discovery of candidate drug targets.
authors.affiliation: GPC Biotech AG, 20 Fraunhoferstrasse, Planegg/Martinsried 82152, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Becker
authors.forename: Krishna
authors.initials: K
authors.lastname: Murthi
authors.forename: Chase
authors.initials: C
authors.lastname: Smith
authors.forename: Jon
authors.initials: J
authors.lastname: Come
authors.forename: Nuria
authors.initials: N
authors.lastname: Costa-Roldán
authors.forename: Christine
authors.initials: C
authors.lastname: Kaufmann
authors.forename: Urs
authors.initials: U
authors.lastname: Hanke
authors.forename: Carsten
authors.initials: C
authors.lastname: Degenhart
authors.forename: Sabine
authors.initials: S
authors.lastname: Baumann
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wallner
authors.forename: Andrea
authors.initials: A
authors.lastname: Huber
authors.forename: Severine
authors.initials: S
authors.lastname: Dedier
authors.forename: Simone
authors.initials: S
authors.lastname: Dill
authors.forename: David
authors.initials: D
authors.lastname: Kinsman
authors.forename: Mark
authors.initials: M
authors.lastname: Hediger
authors.forename: Nicholas
authors.initials: N
authors.lastname: Bockovich
authors.forename: Sebastian
authors.initials: S
authors.lastname: Meier-Ewert
authors.forename: Arthur F
authors.initials: AF
authors.lastname: Kluge
authors.forename: Nikolai
authors.initials: N
authors.lastname: Kley
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D008024:Ligands; D040081:Protein Array Analysis; D020543:Proteome / Q000235:genetics; D012440:Saccharomyces / Q000235:genetics; D020798:Two-Hybrid System Techniques
chemical_list: D004791:Enzyme Inhibitors; D008024:Ligands; D020543:Proteome; C113584:purvalanol B; D016203:CDC2 Protein Kinase; D018844:Cyclin-Dependent Kinases; D000225:Adenine