Yan Kang, Hunt Eric, Berge John, May Earl, Copeland Robert A, Gontarek Richard R
1250 South Collegeville Road, Collegeville, PA 19426, USA.
Antimicrob Agents Chemother. 2005 Aug;49(8):3367-72. doi: 10.1128/AAC.49.8.3367-3372.2005.
A fluorescence polarization assay is described that measures the binding of fluorescently labeled erythromycin to 70S ribosomes from Escherichia coli and the displacement of the erythromycin from these ribosomes. The assay has been validated with several macrolide derivatives and other known antibiotics. We demonstrate that this assay is suitable for determining the dissociation constants of novel compounds that have binding sites overlapping those of macrolides. This homogeneous binding assay provides a valuable tool for defining structure-activity relationships among compounds during the discovery and development of new ribosome-targeting drugs.
本文描述了一种荧光偏振测定法，该方法用于测量荧光标记的红霉素与大肠杆菌70S核糖体的结合以及红霉素从这些核糖体上的解离。该测定法已通过几种大环内酯衍生物和其他已知抗生素进行了验证。我们证明，该测定法适用于确定具有与大环内酯类药物重叠结合位点的新型化合物的解离常数。这种均相结合测定法为在新型核糖体靶向药物的发现和开发过程中定义化合物之间的构效关系提供了一种有价值的工具。
