A general NMR method for rapid, efficient, and reliable biochemical screening

被引:102
作者
Dalvit, C
Ardini, E
Flocco, M
Fogliatto, GP
Mongelli, N
Veronesi, M
机构
[1] Pharmacia, Dept Chem, I-20014 Nerviano, MI, Italy
[2] Pharmacia, Dept Biol, I-20014 Nerviano, MI, Italy
关键词
D O I
10.1021/ja038128e
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
High-throughput screening is usually the method of drug-lead discovery. It is now well accepted that, for a functional assay, quality is more important than quantity. The ligand-based or protein-based NMR screening methodologies for detecting compounds binding to the macromolecular target of interest are now well established. A novel and sensitive NMR method for rapid, efficient, and reliable biochemical screening is presented. The method named 3-FABS (three fluorine atoms for biochemical screening) requires the labeling of the substrate with a CF3 moiety and utilizes F-19 NMR spectroscopy for the detection of the starting and enzymatically modified substrates. The method allows for high-quality screening of large compound or natural product extract collections and for measuring their IC50 values. Applications of this technique to the screening of inhibitors of the Ser/Thr kinase AKT1 and the protease trypsin are presented. In addition, an interesting application of 3-FABS to functional genomics is also presented.
引用
收藏
页码:14620 / 14625
页数:6
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