针对铜绿假单胞菌毒力蛋白伪唾液酸酶Pse的新药设计分子对接研究

Molecular docking of the oseltamivir analogs to the pseudaminidase in Pseudomonas aeruginosa for new inhibitor design

目的揭示铜绿假单胞菌伪神经氨基酸(Pse)耐奥司他韦类似药物的分子机制,开展新药设计分子对接研究,为防治流行性感冒感染后继发性肺炎提供理论基础。方法克隆、表达和提纯重组Pse蛋白;利用蛋白结构分析确定Pse关键活性位点氨基酸,点突变并以酶学方法验证,并采取分子对接技术进行新药前体的平面设计。结果奥司他韦类似药物对铜绿假单胞菌Pse没有抑制作用,Phe129位氨基酸是引起耐药的主要原因,对奥司他韦类似药物C-5支链改造是Pse抑制剂设计的唯一有效途径。结论奥司他韦类似药物对于铜绿假单胞菌Pse不能产生抑制作用,只有进行改造后的伪唾液酸才能对Pse起到抑制作用。

Objective To probe the molecular mechanisms of oseltamivir analog resistance of Pse in Pseudomonas aeruginosa and perform molecular docking for new inhibitor design to provide further insights of pneumonia secondary to influenza infections.Methods Pse was cloned,over-expressed and purified.The key residue confers oseltamivir analog resistance was mapped via threedimensional structure.Site mutation analysis and enzyme assays were performed to confirm the hypothesis.Molecular docking of the oseltamivir analogs to Pse for new inhibitor design.Results Pse from Pseudomonas aeruginosa was found to be fully resistant to oseltamivir analogs.Phe129 handicapped the binding of these drugs.C-5 group modification of oseltamivir analogs could be the only effective way of new inhibitor design.Conclusions Oseltamivir analogs cannot inhibit Pse while modified substrate such as pseudominic acid might do.