WaterLOGSY as a method for primary NMR screening: Practical aspects and range of applicability
WaterLOGSY as a method for primary NMR screening: Practical aspects and range of applicability
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DOI：
10.1023/a:1013302231549
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发表时间：
2001-12-01
影响因子：
2.7
通讯作者：
Stockman, B
中科院分区：
文献类型：
--
作者：
Dalvit, C;Fogliatto, G;Stockman, B
WaterLOGSY 代表了一种强大的初级 NMR 筛选方法，用于鉴定与大分子（包括蛋白质和 DNA 或 RNA 片段）相互作用的化合物。实验中建设性地使用了几种中继途径，将大量水磁化强度转移到配体。该方法对于鉴定微摩尔亲和力的新型支架特别有用，然后可以使用定向筛选、组合化学、药物化学和基于结构的药物设计对其进行优化。这里详细分析了 WaterLOGSY 实验的实际方面和适用范围。竞争结合和滴定 WaterLOGSY 允许在适当校正后评估解离结合常数。该技术的高灵敏度与用于识别活性成分的混合物的简单解卷积相结合，显着减少了 NMR 筛选过程所需的材料量和时间。
WaterLOGSY represents a powerful method for primary NMR screening in the identification of compounds interacting with macromolecules, including proteins and DNA or RNA fragments. Several relay pathways are used constructively in the experiment for transferring bulk water magnetization to the ligand. The method is particularly useful for the identification of novel scaffolds of micromolar affinity that can be then optimized using directed screening, combinatorial chemistry, medicinal chemistry and structure-based drug design. The practical aspects and range of applicability of the WaterLOGSY experiment are analyzed in detail here. Competition binding and titration WaterLOGSY permit, after proper correction, the evaluation of the dissociation binding constant. The high sensitivity of the technique in combination with the easy deconvolution of the mixtures for the identification of the active components, significantly reduces the amount of material and time needed for the NMR screening process.