Automated Protein-Ligand Interaction Screening by Mass Spectrometry

作者：Maple Hannah J; Garlish Rachel A; Rigau Roca Laura; Porter John; Whitcombe Ian; Prosser Christine E; Kennedy Jeff; Henry Alistair J; Taylor Richard J; Crump Matthew P^*; Cro**y John

来源：Journal of Medicinal Chemistry, 2012, 55(2): 837-851.

DOI：10.1021/jm201347k

Identifying protein-ligand binding interactions is a key step during early-stage drug discovery. Existing screening techniques are often associated with drawbacks such as low throughput, high sample consumption, and dynamic range limitations. The increasing use of fragment-based drug discovery (FBDD) demands that these techniques also detect very weak interactions (mM K-D values). This paper presents the development and validation of a fully automated screen by mass spectrometry, capable of detecting fragment binding into the millimolar K-D range. Low sample consumption, high throughput, and wide dynamic range make this a highly attractive, orthogonal approach. The method was applied to screen 157 compounds in 6 h against the anti-apoptotic protein target Bcl-x(L). Mass spectrometry results were validated using STD-NMR, HSQC-NMR, and ITC experiments. Agreement between techniques suggests that mass spectrometry offers a powerful, complementary approach for screening.

• 出版日期2012-1-26