An ultraefficient affinity-based high-throughout screening process: Application to bacterial cell wall biosynthesis enzyme MurF
An ultraefficient affinity-based high-throughout screening process: Application to bacterial cell wall biosynthesis enzyme MurF
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DOI：
10.1177/1087057106289971
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发表时间：
2006-10-01
影响因子：
--
通讯作者：
Beutel, Bruce A.
中科院分区：
文献类型：
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作者：
Comess, Kenneth M.;Schurdak, Mark E.;Beutel, Bruce A.
作者描述了通过一种新的亲和筛选方法发现了一类新的肺炎链球菌细胞壁生物合成酶MurF的抑制剂。该策略涉及筛选非常大的混合物的不同的小有机分子对蛋白质目标的基础上的平衡结合，然后通过迭代的超滤步骤和质谱配体鉴定。来自任何基于亲和力的筛选方法的命中通常可以是相对非选择性的配体，有时被称为“滋扰”或“混杂”化合物。通过对文库中经验确定的混杂化合物的子集进行电子减法，容易地鉴定出对MurF靶具有结合亲和力选择性的配体，而无需随后的选择性小组。发现和鉴定新的特异性配体的完整策略可以应用于所有可溶性蛋白质靶标和各种各样的配体文库。
The authors describe the discovery of a new class of inhibitors to an essential Streptococcus pneumoniae cell wall biosynthesis enzyme, MurF, by a novel affinity screening method. The strategy involved screening very large mixtures of diverse small organic molecules against the protein target on the basis of equilibrium binding, followed by iterative ultrafiltration steps and ligand identification by mass spectrometry. Hits from any affinity-based screening method often can be relatively nonselective ligands, sometimes referred to as "nuisance" or "promiscuous" compounds. Ligands selective in their binding affinity for the MurF target were readily identified through electronic subtraction of an empirically determined subset of promiscuous compounds in the library without subsequent selectivity panels. The complete strategy for discovery and identification of novel specific ligands can be applied to all soluble protein targets and a wide variety of ligand libraries.