Chemical screening by mass spectrometry to identify inhibitors of anthrax lethal factor
Chemical screening by mass spectrometry to identify inhibitors of anthrax lethal factor
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DOI：
10.1038/nbt973
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发表时间：
2004-06-01
影响因子：
46.9
通讯作者：
Mrksich, M
中科院分区：
文献类型：
--
作者：
Min, DH;Tang, WJ;Mrksich, M
质谱（MS）分析适用于广泛的生物分析物，并且具有不需要标记分析物的重要优点。然而，MS方法的一个缺点是需要色谱步骤来制备分析物，这使得MS无法用于化学筛选和快速分析。在这里，我们报告说，表面化学定制的表征基质辅助激光解吸电离飞行时间MS消除了样品处理的需要，使这种技术适用于并行筛选实验。定制的基底是基于自组装单层，其呈现与靶蛋白和酶相互作用的配体。我们应用这种方法来筛选针对炭疽致死因子的蛋白酶活性的化学文库，并报道了一种化合物，其抑制致死因子活性的Ki为1.1 μ M，并阻断293细胞中MEK 1的切割。
Mass spectrometry (MS) analysis is applicable to a broad range of biological analytes and has the important advantage that it does not require analytes to be labeled. A drawback of MS methods, however, is the need for chromatographic steps to prepare the analyte, precluding MS from being used in chemical screening and rapid analysis. Here, we report that surfaces that are chemically tailored for characterization by matrix-assisted laser-desorption ionization time-of-flight MS eliminate the need for sample processing and make this technique adaptable to parallel screening experiments. The tailored substrates are based on self-assembled monolayers that present ligands that interact with target proteins and enzymes. We apply this method to screen a chemical library against protease activity of anthrax lethal factor, and report a compound that inhibits lethal factor activity with a K-i of 1.1 muM and blocks the cleavage of MEK1 in 293 cells.