关于新耐药蛋白OptrA的2个关键活性位点  

Confirmation of two key active sites of the new drug resistance protein OptrA

作　　者：钟晓波[1] 王琳[1] 王铁东[1] 钟灵[1] 王大成[1] 

机构地区：[1]吉林大学动物科学学院,吉林长春130062

出　　处：《中国兽医学报》2017年第4期717-720,共4页Chinese Journal of Veterinary Science

基　　金："十三五"国家重点研发计划资助项目(2016YFD05013)

摘　　要：通过ATP水解酶活性试验分析2个突变位点对OptrA活性的影响,并通过最低抑菌浓度试验分析不同突变位点对金黄色葡萄球菌耐药性的影响。将OptrA蛋白作抗原免疫日本大耳白兔,提取血清抗体,用Western bolt分析突变前后蛋白表达量的变化。结果显示,当把全长氨基酸序列208和488位的谷氨酸(E)突变后,OptrA在金黄色葡萄球菌中的表达量不变,但其ATP水解活性分别降低30%和70%,并且都导致金黄色葡萄球菌失去了对氟苯尼考的耐药性。说明OptrA需要水解ATP才能介导细菌耐药性,为以后研究新耐药蛋白OptrA的抑制剂和肠球菌感染的防制奠定基础。The effects of two mutation sites on OptrA ATP hydrolase were analyzed by ATPase activity assay,and the effect of different mutation sites on drug resistance of Staphylococcus aureus was analyzed by the minimum inhibitory concentration experiment. Japanese white rabbits immuned with OptrA protein was used to make serum antibody. Western blot analysis was used to analyze the changes of protein expression before and after the mutation. Results showed that when the glutamate at the 208 and 488 positions of full length amino acid sequence were mutated, the expression of OptrA unchanged,but its ATP hydrolysis activity were reduced by 30% and 70% ,and led to Staphylococcus aureus lost the resistance to florfenicol. This indicated that OptrA needed to hydrolyze ATP in mediating bacterial resistance,thus laying the groundwork for the future study on inhibitors of the new resistance protein OptrA and prevention of enterococcal infections.

关 键 词：OptrA ATP水解酶 耐药性 

分 类 号：S852.61[农业科学—基础兽医学]