畜牧与兽医2024，Vol.56Issue(3)：31-38,8.
穿心莲内酯对耐甲氧西林金黄色葡萄球菌关键耐药蛋白PBP2a的抑制作用
Inhibition of key resistance protein PBP2a of methicillin-resistant Staphylococcus aureus by andrographolide
摘要
Abstract
This study was to explore the inhibitory effect of andrographolide(AP)on the critical drug-resistant and protein penicillin-binding protein 2a(PBP2a)of methicillin-resistant Staphylococcus aureus(MRSA).Here,the effect of AP on mecA gene transcription was detected by real-time fluorescence quantitative PCR.The protein structure was analyzed by the online software SWISS-MODEL.Pymol and AutoDock Tools software performed molecular docking to explore the potential mechanism of AP binding to PBP2a,and kinetic simulation was used to verify the reliability of AP binding to PBP2a protein.Then,prokaryotic expression of PBP2a was further performed,and its poly-clonal antibody was prepared.The change of PBP2a protein content in MRSA under the action of AP was detected by Western blot.The re-sults showed that AP significantly down-regulated the transcription level of mecA at 64 μg/mL,and a stable hydrogen bond force was formed between AP and GLU170,GLU239,and THR238 of PBP2a protein.The transpeptidase domain of 38 kDa PBP2a protein was successfully expressed and purified,and the prepared polyclonal antibody specifically bound to the PBP2a protein.With the increase in AP concentration,the expression of PBP2a decreased gradually.In conclusion,the molecular simulation in this study suggested that AP had the potential to in-hibit PBP2a protein.The present results indicated that AP could inhibit the expression of PBP2a by inhibiting the transcription of mecA,thus revealing the possible mechanism of AP sensitizing MRSA to β-lactam antibiotics.关键词
耐甲氧西林金黄色葡萄球菌/PBP2a/穿心莲内酯/分子模拟Key words
methicillin-resistant Staphylococcus aureus/PBP2a/andrographolide/molecular simulation分类
农业科技引用本文复制引用
李彩霞,刘盼盼,陈晓慧,罗小凤,王桂琴..穿心莲内酯对耐甲氧西林金黄色葡萄球菌关键耐药蛋白PBP2a的抑制作用[J].畜牧与兽医,2024,56(3):31-38,8.基金项目
宁夏自然科学基金重点项目(No.2020AAC02009) （No.2020AAC02009）
