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 bond 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.
Original language | English (US) |
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Pages (from-to) | 370-375 |
Number of pages | 6 |
Journal | MedChemComm |
Volume | 5 |
Issue number | 3 |
DOIs | |
State | Published - Mar 2014 |