High-Throughput Screening (HTS) by NMR Guided Identification of Novel Agents Targeting the Protein Docking Domain of YopH.
High-Throughput Screening (HTS) by NMR Guided Identification of Novel Agents Targeting the Protein Docking Domain of YopH.
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DOI：
10.1002/cmdc.201500441
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发表时间：
2016-04-19
期刊：
影响因子：
3.4
通讯作者：
Pellecchia M
中科院分区：
文献类型：
--
作者：
Bottini A;Wu B;Barile E;De SK;Leone M;Pellecchia M
最近，我们描述了一种新方法，称为 HTS by NMR，它允许从大型组合肽库中识别针对给定靶标的有效且选择性的肽模拟物。在这里，我们采用 NMR 方法的 HTS 来设计针对耶尔森氏菌外部蛋白 H (YopH-NT) 氨基末端结构域的新型类肽序列。我们的目的是破坏 YopH-NT 与其细胞底物之间的蛋白质-蛋白质相互作用，从而间接抑制 YopH 酶功能。这些研究产生了一种序列为 Ac-F-pY-cPG-D-P-NH2（pY = 磷酸酪氨酸；cPG = 环戊基甘氨酸）的新型试剂，其针对 YopH-NT 的 Kd 值为 310 nM。我们证明了 YopH-NT 的这种药理学抑制剂导致全长 YopH 抑制细胞底物的去磷酸化。因此，这种药物可能成为开发针对耶尔森氏菌感染的新型疗法的宝贵垫脚石。报告的数据进一步证明了 HTS 通过 NMR 方法在衍生针对蛋白质-蛋白质相互作用的新型肽模拟物方面的效用。
Recently we described a novel approach, named HTS by NMR that allows the identification, from large combinatorial peptide libraries, of potent and selective peptide mimetics against a given target. Here we deployed the HTS by NMR approach for the design of novel peptoid sequences targeting the amino terminal domain of the Yersinia outer protein H (YopH-NT). We aimed at disrupting the protein-protein interactions between YopH-NT and its cellular substrates, with the goal of inhibiting indirectly YopH enzymatic function. These studies resulted in a novel agent of sequence Ac-F-pY-cPG-D-P-NH2 (pY = phosphotyrosine; cPG = cyclopentyl glycine) with a Kd value against YopH-NT of 310 nM. We demonstrated that such pharmacological inhibitor of YopH-NT resulted in the inhibition of the dephosphorylation of a cellular substrate by full length YopH. Hence, potentially this agent represents a valuable stepping stone for the development of novel therapeutics against Yersinia infections. The data reported further demonstrate the utility of the HTS by NMR approach in deriving novel peptide-mimetics targeting protein-protein interactions.