Fluorine-NMR competition binding experiments for high-throughput screening of large compound mixtures
Fluorine-NMR competition binding experiments for high-throughput screening of large compound mixtures
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DOI：
10.2174/1386207023329923
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发表时间：
2002-12-01
影响因子：
1.8
通讯作者：
Stockman, BJ
中科院分区：
文献类型：
--
作者：
Dalvit, C;Flocco, M;Stockman, BJ
基于高通量配体的 NMR 筛选和竞争结合实验已扩展到 F-19 检测。氟是这些实验的有利核，因为当与大分子靶标结合时，化学位移各向异性 (CSA) 对配体信号的 F-19 横向弛豫有显着贡献。含有氟原子的低至中等亲和力配体用作检测和表征新配体的参考分子。使用选定的参考化合物进行滴定 NMR 实验，以找到 HTS 的最佳设置条件并推导已识别的 NMR 命中的结合常数。由于 F-19 核的高灵敏度以及与待筛选混合物信号不存在重叠，因此可以对大型化学混合物和植物或真菌提取物针对感兴趣的受体进行快速 HTS。最后，提出了一种使用参考分子与对照分子相结合的 HTS 的新方法。
High-throughput ligand-based NMR screening with competition binding experiments is extended to F-19 detection. Fluorine is a favorable nucleus for these experiments because of the significant contribution of the Chemical Shift Anisotropy (CSA) to the F-19 transverse relaxation of the ligand signal when bound to a macromolecular target. A low to moderate affinity ligand containing a fluorine atom is used as a reference molecule for the detection and characterization of new ligands. Titration NMR experiments with the selected reference compound are performed for finding the optimal set-up conditions for HTS and for deriving the binding constants of the identified NMR hits. Rapid HTS of large chemical mixtures and plant or fungi extracts against the receptor of interest is possible due to the high sensitivity of the F-19 nucleus and the absence of overlap with the signals of the mixtures to be screened. Finally, a novel approach for HTS using a reference molecule in combination with a control molecule is presented.