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Medchemcomm. 2014 Mar 1;5:370-375.

FP Tethering: a screening technique to rapidly identify compounds that disrupt protein-protein interactions.

Author information

1
Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States ; Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109, United States.
2
Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94158, United States.
3
Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, United States.
4
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.

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.

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