FP tethering: a screening technique to rapidly identify compounds that disrupt protein–protein interactions
| Autor: | T. Justin Rettenmaier, Anna K. Mapp, Jean M. Lodge, William C. K. Pomerantz, James A. Wells |
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| Rok vydání: | 2014 |
| Předmět: |
Pharmacology
chemistry.chemical_classification Chemistry Tethering Organic Chemistry Pharmaceutical Science Nanotechnology Peptide Biochemistry Article Disulphide bond formation Protein–protein interaction Transcriptional Coactivator Drug Discovery Biophysics Molecular Medicine Protein target Peptide ligand Fluorescence anisotropy |
| Zdroj: | MedChemComm. 5:370-375 |
| ISSN: | 2040-2511 2040-2503 |
| DOI: | 10.1039/c3md00356f |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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