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维吉尼亚霉素乙酰转移酶D(VatD)通过灭活链阳霉素A而在链阳霉素耐药性的产生中起重要作用。本研究采用虚拟筛选技术寻找VatD的抑制剂,此VatD抑制剂可以和链阳霉素联合使用,从而提供新的治疗耐药菌感染的方法。作者首次应用基于结构的虚拟筛选方法(分子对接)从含300000化合物的商业化数据库中筛选对抗VatD底物结合位点的化合物,从200个评分最高的化合物中选取26个测定对VatD酶活性的抑制作用。将构建的质粒pRSETB/vatD转染宿主细胞E.coli(TrxB)用于过表达,纯化的VatD对维吉尼亚霉素M1表现乙酰转移酶活性。26个化合物中有3个对VatD表现抑制作用,IC50分别为168.6,91.0和55.2μmol.L-1。其他化合物在反应体系中不溶解和/或对酶活性的抑制作用很小(IC50>200μmol.L-1)。本文首次设计VatD的小分子化合物抑制剂,发现了3个有活性的化合物,希望其可作为先导化合物进一步发展为新的对抗链阳霉素耐药性的药物。
Virulentmycin acetyltransferase D (VatD) plays an important role in the production of streptomycin resistance by inactivating streptogramin. In this study, a virtual screening technique was used to find VatD inhibitors. This VatD inhibitor can be used in combination with streptomycin so as to provide a new method for the treatment of drug-resistant bacterial infections. For the first time, the authors used a structural-based virtual screening approach (molecular docking) to screen compounds against VatD substrate binding sites from commercial databases containing 300,000 compounds and select 26 of the 200 most-rated compounds for their ability to detect VatD activity Inhibition. The constructed plasmid pRSETB / vatD was transfected into host cell E. coli (TrxB) for overexpression, purified VatD to demonstrate acetyltransferase activity on virginiamycin M1. Three out of 26 compounds showed an inhibitory effect on VatD with IC50 of 168.6, 91.0 and 55.2 μmol.L-1, respectively. Other compounds did not dissolve in the reaction system and / or had little inhibitory effect on the enzyme activity (IC50> 200 μmol.L-1). In this paper, for the first time, VatD, a small molecule compound inhibitor, was designed and three active compounds were found in the hope of further developing them into lead compounds against the drug resistance of cingomycin.