Insight into the Dual inhibitory Mechanism of verbascoside targeting serine/threonine phosphatase Stp1 against Staphylococcus aureus.
Insight into the Dual inhibitory Mechanism of verbascoside targeting serine/threonine phosphatase Stp1 against Staphylococcus aureus.
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DOI：
10.1016/j.ejps.2020.105628
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发表时间：
2020-10
期刊：
影响因子：
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通讯作者：
Yanan Yang;Xiyan Wang;Yawen Gao;Hongsu Wang;X. Niu
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文献类型：
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作者：
Yanan Yang;Xiyan Wang;Yawen Gao;Hongsu Wang;X. Niu
类真核丝氨酸/苏氨酸磷酸酶（Stp1）是一种酶依赖性蛋白磷酸酶，参与调节金黄色葡萄球菌的多种毒力因子。由于其在 S 中的作用。在金黄色葡萄球菌感染中，Stp1已成为抗生素开发的潜在靶点。不幸的是，描述 Stp1 抑制剂的报道很少。通过虚拟筛选，我们发现了一种强效且有效的 Stp1 抑制剂——毛蕊花苷 (VBS)。有趣的是，酶反应的动力学表明，这种天然抑制剂通过竞争和变构机制发挥作用。为了探索 VBS 和 Stp1 之间的相互作用机制，对 Stp1-VBS 复合物进行了标准分子动力学 (MD) 模拟。与实验结果一致，在 Stp1 中鉴定出了 VBS 的竞争性和变构结合位点。 Met39、Gly41、His42、Arg161 和 Asn162 残基参与 VBS 的竞争性结合，而 Arg122、Ser136、Asp137、Asn142 和 Val145 残基与 VBS 的变构结合相关。这些残基的贡献通过氨基酸定点诱变和荧光猝灭实验得到证实。这项工作表明，VBS 是一种有效的抗金黄色葡萄球菌毒力化合物。金黄色葡萄球菌感染，为进一步开发新型抗毒剂奠定了基础。
The eukaryotic-like serine/threonine phosphatase (Stp1) is an enzyme-dependent protein phosphatase involved in regulating various virulence factors of Staphylococcus aureus. Owing to its role inS. aureusinfections, Stp1 has become a potential target for antibiotic development. Unfortunately, there are very few reports describing Stp1 inhibitors. Using virtual screening, we have identified a potent and effective Stp1 inhibitor, verbascoside (VBS). Interestingly, the kinetics of the enzymatic reaction revealed that this natural inhibitor acts via both competitive and allosteric mechanisms. To explore the mechanism of interaction between VBS and Stp1, standard molecular dynamics (MD) simulations were performed for the Stp1-VBS complex. Consistent with the experimental results, competitive and allosteric binding sites for VBS were identified in Stp1. Met39, Gly41, His42, Arg161, and Asn162 residues were involved in the competitive binding of VBS, while Arg122, Ser136, Asp137, Asn142, and Val145 residues were associated with the allosteric binding of VBS. The contributions of these residues were confirmed by amino acid site-directed mutagenesis and fluorescence quenching experiments. This work demonstrates that VBS is a potent anti-virulence compound againstS. aureusinfection, laying the foundation for the further development of novel anti-virulence agents.