核糖核酸酶A超家族抗菌活性评价  

An Evaluation of the Antimicrobial Activity of the Ribonuclease A Superfamily

作　　者：朱泽浩 励航宇 孙钧 盛静浩 ZHU Ze-Hao;LI Hang-Yu;SUN Jun;SHENG Jing-Hao(Affiliated Hangzhou First People’s Hospital,Zhejiang University School of Medicine,Hangzhou 310006,China;Institute of Environmental Medicine,Zhejiang University School of Medicine,Hangzhou 310058,China;College of Life Science,Zhejiang University,Hangzhou 30058,China)

机构地区：[1]浙江大学医学院附属杭州市第一人民医院,杭州310006 [2]浙江大学医学院环境医学研究所,杭州310058 [3]浙江大学生命科学学院,杭州310058

出　　处：《中国生物化学与分子生物学报》2023年第4期524-530,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金联合基金(No.U21A20202)和国家自然科学基金面上项目(No.32071289)资助。

摘　　要：抗菌肽是机体重要的免疫防御分子,具有广谱杀菌活性。核糖核酸酶A是脊椎动物特异性的分泌型蛋白质,在人基因组中包含8个经典成员(RNase1-8)。它们作为一类重要的抗菌肽,广泛分布于机体需要抵抗外界病原微生物的组织中,除了特有的生物学功能外,均具有一定的抗菌活性。然而,目前各实验室对它们抗菌活性的报道并不一致,有必要开展横向比较分析。为此,我们表达纯化了人核糖核酸酶A超家族8个成员的重组蛋白质,并在同一实验条件下以半致死浓度评估了它们对革兰氏阴性菌(大肠杆菌)和革兰氏阳性菌(金黄色葡萄球菌)的抗菌活性。结果显示,RNase1-8重组蛋白质对大肠杆菌半数致死浓度分别为:0.081,0.046,0.008,0.250,2.028,0.072,0.001μmol·L~(-1)和1.1416μmol·L~(-1);对金黄色葡萄球菌半数致死浓度分别为:3.427,1.856,2.211,5.188,8.274,4.356,2.502μmol·L~(-1)和9.916μmol·L~(-1)。该结果提示,RNase1-8对大肠杆菌的抗菌活性存在明显差异,其中RNase3和RNase7活性最强,且均显著高于各自对金黄色葡萄球菌的抗菌活性;8个成员对金黄色葡萄球菌的抗菌活性无明显差异。据此我们认为,核糖核酸酶A超家族对革兰氏阴性菌具有较好的杀伤活性,可用于革兰氏阴性菌抗菌产品的开发。Antimicrobial peptides,which hold a broad spectrum of bactericidal effects,are important immune defense molecules of the human body.Ribonuclease A is a vertebrate-specific secretory protein that has eight members(RNase1-8)in the human proteome.As an important class of antimicrobial peptides,they are widely distributed in the tissues that need to resist pathogenic microorganisms,and all of them have certain antimicrobial activities in addition to their own unique biological functions.However,reports on their antibacterial activities are not consistent from various laboratories,thus a cross-sectional comparative analysis is needed.To this end,we expressed and purified recombinant proteins of eight classical members of the human ribonuclease A superfamily,and evaluated their antibacterial activities against Gram-negative(Escherichia coli)and Gram-positive(Staphylococcus aureus)bacteria with 50%lethal concentrations under the same experimental condition.The results showed that the 50%lethal concentrations of recombinant RNase1-8 proteins against Escherichia coli were 0.0811,0.046,0.008,0.250,2.028,0.072,0.001μmol·L-1 and 1.1416μmol·L-1,respectively;that against Staphylococcus aureus were 3.427,1.856,2.211,5.188,8.274,4.356,2.502μmol·L-1 and 9.916μmol·L-1,respectively.The results indicate that there is a significant difference in RNase1-8 antibacterial activities against Escherichia coli,among which RNase3 and RNase7 were most active,and both of them had significantly higher antibacterial activities than their activities against Staphylococcus aureus.Meanwhile,there were no significant differences in their antibacterial activity against Staphylococcus aureus.Therefore,we conclude that the Ribonuclease A superfamily has a higher killing activity against Gram-negative bacteria,and may have the potential to be further developed as antibacterial products.

关 键 词：核糖核酸酶A 抗菌肽 大肠杆菌 金黄色葡萄球菌 抗菌活性 

分 类 号：Q51[生物学—生物化学]