In this article we describe the production and screening of a genetically encoded library of 10⁶ lanthipeptides in Escherichia coli using the substrate-tolerant lanthipeptide synthetase ProcM. This plasmid-encoded library was combined with a bacterial reverse two-hybrid system for the interaction of the HIV p6 protein with the UEV domain of the human TSG101 protein, which is a critical protein–protein interaction for HIV budding from infected cells. Using this approach, we identified an inhibitor of this interaction from the lanthipeptide library, whose activity was verified in vitro and in cell-based virus-like particle-budding assays. Given the variety of lanthipeptide backbone scaffolds that may be produced with ProcM, this method may be used for the generation of genetically encoded libraries of natural product–like lanthipeptides containing substantial structural diversity. Such libraries may be combined with any cell-based assay to identify lanthipeptides with new biological activities.

在本文中，我们描述了使用底物耐受的羊毛硫肽合成酶 ProcM 在大肠杆菌中产生和筛选 10 ^{6 种}羊毛硫肽的基因编码文库。该质粒编码文库与细菌反向双杂交系统相结合，用于 HIV p6 蛋白与人类 TSG101 蛋白的 UEV 结构域的相互作用，这是 HIV 从受感染细胞中出芽的关键蛋白质-蛋白质相互作用。使用这种方法，我们从羊毛硫肽库中鉴定出了这种相互作用的抑制剂，其活性在体外和基于细胞的病毒样颗粒出芽测定中得到了验证。鉴于 ProcM 可以生产多种羊毛硫肽骨架支架，该方法可用于生成含有大量结构多样性的天然产物样羊毛硫肽的基因编码文库。此类文库可以与任何基于细胞的测定法结合以鉴定具有新生物活性的羊毛硫肽。