Author/Authors :
Frank Becker، نويسنده , , Krishna Murthi، نويسنده , , Chase Smith، نويسنده , , Jon Come، نويسنده , , Nuria Costa-Rold?n، نويسنده , , Christine Kaufmann، نويسنده , , Urs Hanke، نويسنده , , Carsten Degenhart، نويسنده , , Sabine Baumann، نويسنده , , Wolfgang Wallner، نويسنده , , Andrea Huber، نويسنده , , Severine Dedier، نويسنده , , Simone Dill، نويسنده , , David Kinsman، نويسنده , , Mark Hediger، نويسنده , , Nicholas Bockovich، نويسنده , , Sebas، نويسنده ,
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.
