FP tethering: a screening technique to rapidly identify compounds that disrupt protein-protein interactions
FP tethering: a screening technique to rapidly identify compounds that disrupt protein-protein interactions
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DOI：
10.1039/c3md00356f
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发表时间：
2014-03-01
期刊：
影响因子：
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通讯作者：
Mapp, Anna K.
中科院分区：
文献类型：
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作者：
Lodge, Jean M.;Rettenmaier, T. Justin;Mapp, Anna K.
系链是一种筛选技术，用于发现通过形成二硫键与预定位点结合的小分子片段。栓系筛选传统上依赖于质谱法来检测二硫键形成，这需要耗时的液相色谱步骤。在这里，我们表明，拴系可以快速，廉价地使用同质荧光偏振（FP）检测位移的肽配体从蛋白质靶作为二硫化物形成的间接读数。我们应用这种方法，称为FP拴系，以确定片段，破坏转录辅激活因子CBP和转录激活肽pKID的KIX结构域之间的蛋白质-蛋白质相互作用。
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.