As resistance to antibiotics increases, the exploration of new targets and strategies to combat pathogenic bacteria becomes more urgent. Ideal protein targets are required for viability across many species, are unique to prokaryotes to limit effects on the host, and have robust assays to quantitate activity and identify inhibitors. Lipoprotein signal peptidase (Lsp) is a transmembrane aspartyl protease required for lipoprotein maturation and comprehensively fits these criteria. Here, we have developed the firstin vitrohigh-throughput assay to monitor proteolysis by Lsp. We employed our high-throughput screen assay against 646,275 compounds to discover inhibitors of Lsp and synthesized a range of analogs to generate molecules with nanomolar half maximal inhibitory concentration values. Importantly, our inhibitors are effective in preventing the growth ofE. colicultures in the presence of outer-membrane permeabilizer PMBN and should facilitate development of antibacterial agents with a novel mechanism of action to treat antibiotic-resistant bacteria.

中文翻译：

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脂蛋白信号肽酶抑制剂具有抗生素特性的鲁棒性体外HT平台设计已被鉴定

随着对抗生素的抗性增加，对抗病原细菌的新靶标和策略的探索变得更加紧迫。理想的蛋白质靶标对于许多物种的生存力都是必需的，是原核生物特有的，以限制对宿主的作用，并具有可靠的测定方法来定量活性和鉴定抑制剂。脂蛋白信号肽酶（Lsp）是脂蛋白成熟所需的跨膜天冬氨酰蛋白酶，完全符合这些标准。在这里，我们开发了第一个体外高通量检测方法来监测Lsp的蛋白水解作用。我们针对646,275种化合物进行了高通量筛选分析，以发现Lsp抑制剂，并合成了一系列类似物以产生具有纳摩尔半数最大抑制浓度值的分子。重要的，我们的抑制剂可有效预防E的生长。在外膜通透性PMBN存在的情况下进行大肠埃希菌培养，应促进抗菌素的开发，并以新颖的作用机制来治疗抗药性细菌。