Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5 E (PO Box 56), 00014, Helsinki, Finland.
University of Ljubljana, Faculty of Pharmacy, Aškerčeva 7, 1000, Ljubljana, Slovenia.
Chembiochem. 2020 Jul 1;21(13):1918-1922. doi: 10.1002/cbic.201900773. Epub 2020 Mar 3.
Quorum sensing (QS), a bacterial communication strategy, has been recognized as one of the control mechanisms of virulence in bacteria. Thus, targeting QS offers an interesting opportunity to impair bacterial pathogenicity and develop antivirulence agents. Aiming to enhance the discovery of QS inhibitors, we developed a bioreporter Escherichia coli JW5505 pET-Plsrlux and set up a cell-based assay for identifying inhibitors of autoinducer-2 (AI-2)-mediated QS. A comparative study on the performance of target- versus cell-based assays was performed, and 91 compounds selected with the potential to target the ATP binding pocket of LsrK, a key enzyme in AI-2 processing, were tested in an LsrK inhibition assay, providing 36 hits. The same set of compounds was tested by the AI-2-mediated QS interference assay, resulting in 24 active compounds. Among those, six were also found to be active against LsrK, whereas 18 might target other components of the pathway. Thus, this AI-2-mediated QS interference cell-based assay is an effective tool for complementing target-based assays, yet also stands as an independent assay for primary screening.
群体感应 (QS) 是一种细菌通讯策略，已被认为是细菌毒力控制机制之一。因此，靶向 QS 为削弱细菌致病性和开发抗毒力药物提供了一个有趣的机会。为了增强对 QS 抑制剂的发现，我们开发了一种生物报告大肠杆菌 JW5505 pET-Plsrlux，并建立了一种基于细胞的测定方法，用于鉴定 AI-2 介导的 QS 抑制剂。对基于靶点和基于细胞的测定方法的性能进行了比较研究，并用该方法对 91 种可能靶向 LsrK （AI-2 处理的关键酶）ATP 结合口袋的化合物进行了测试，得到了 36 个阳性结果。用 AI-2 介导的 QS 干扰测定法对同一组化合物进行了测试，得到了 24 个活性化合物。其中，有 6 种对 LsrK 也具有活性，而 18 种可能靶向该途径的其他成分。因此，这种基于 AI-2 的 QS 干扰细胞测定法是对基于靶点的测定法的有效补充，但也可作为初步筛选的独立测定法。
