title: FP Tethering: a screening technique to rapidly identify compounds that disrupt protein-protein interactions.
abstract: Tethering is a screening technique for discovering small-molecule fragments that bind to pre-determined sites via formation of a disulphide bond. Tethering screens traditionally rely upon mass spectrometry to detect disulphide bind formation, which requires a time-consuming liquid chromatography step. Here we show that Tethering can be performed rapidly and inexpensively using a homogenous fluorescence polarization (FP) assay that detects displacement of a peptide ligand from the protein target as an indirect readout of disulphide formation. We apply this method, termed FP Tethering, to identify fragments that disrupt the protein-protein interaction between the KIX domain of the transcriptional coactivator CBP and the transcriptional activator peptide pKID.
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States ; Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109, United States.
authors.forename: Jean M
authors.initials: JM
authors.lastname: Lodge
authors.affiliation: Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94158, United States.
authors.forename: T Justin
authors.initials: TJ
authors.lastname: Rettenmaier
authors.affiliation: Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94158, United States.
authors.forename: James A
authors.initials: JA
authors.lastname: Wells
authors.affiliation: Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, United States.
authors.forename: William C
authors.initials: WC
authors.lastname: Pomerantz
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States ; Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109, United States ; Program in Chemical Biology, University of Michigan, Ann Arbor, Michigan 48109, United States.
authors.forename: Anna K
authors.initials: AK
authors.lastname: Mapp