A General NMR Method for Rapid, Efficient, and Reliable Biochemical Screening

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• 作者：Claudio Dalvit ; Elena Ardini ; Maria Flocco ; Gian Paolo Fogliatto ; Nicola Mongelli ; and Marina Veronesi
• 刊名：Journal of the American Chemical Society
• 出版年：2003
• 出版时间：November 26, 2003
• 年：2003
• 卷：125
• 期：47
• 页码：14620 - 14625
• 全文大小：91K
• 年卷期：v.125,no.47(November 26, 2003)
• ISSN：1520-5126

文摘

High-throughput screening is usually the method of drug-lead discovery. It is now well acceptedthat, for a functional assay, quality is more important than quantity. The ligand-based or protein-basedNMR screening methodologies for detecting compounds binding to the macromolecular target of interestare now well established. A novel and sensitive NMR method for rapid, efficient, and reliable biochemicalscreening is presented. The method named 3-FABS (three fluorine atoms for biochemical screening) requiresthe labeling of the substrate with a CF₃ moiety and utilizes ¹⁹F NMR spectroscopy for the detection of thestarting and enzymatically modified substrates. The method allows for high-quality screening of largecompound or natural product extract collections and for measuring their IC₅₀ values. Applications of thistechnique 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.