抗菌肽BuforinⅡ衍生物特异性结合金黄色葡萄球菌DNA抑菌机理的研究

doi:10.11843/j.issn.0366-6964.2014.07.018

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (7): 1154-1161.doi: 10.11843/j.issn.0366-6964.2014.07.018

抗菌肽BuforinⅡ衍生物特异性结合金黄色葡萄球菌DNA抑菌机理的研究

郝刚^1*，乐国伟²，施用晖²

（1. 西南民族大学生命科学与技术学院，成都 610041；2．江南大学食品学院，无锡 214122）

收稿日期:2014-01-12 出版日期:2014-07-23 发布日期:2014-07-23
通讯作者: 郝刚
作者简介:郝刚（1978-），男，重庆人，博士，主要从事抗菌肽的研究，Tel：86-28-88434002；E-mail：indianahg@hotmail.com
基金资助:
中央高校基本科研业务费（2014NZYQN46）；国家自然科学基金（31172214）；十二五支撑计划（2012BAD33B05）

The Bactericidal Mechanism of Two Analogues of the Antimicrobial Peptide BuforinⅡSpecific Binding to Staphylococcus aureus Genomic DNA

HAO Gang^1*，LE Guo-wei²，SHI Yong-hui²

(1.College of Life Science and Technology， Southwest University for Nationalities， Chengdu 610041， China;2. School of Food Science and Technology， Jiangnan University， Wuxi 214122， China)

Received:2014-01-12 Online:2014-07-23 Published:2014-07-23

摘要/Abstract

摘要：

旨在研究抗菌肽Buforin Ⅱ的衍生物BF2-A/B作用于金黄色葡萄球菌基因组DNA的抑菌机制。原子力显微镜观察了BF2-A/B对金黄色葡萄球菌基因组DNA的结合作用，用凝胶阻滞试验分析了BF2-A/B结合DNA的能力，DNaseⅠ足迹试验研究衍生肽与基因组DNA结合的特异性，并考察了对细胞呼吸作用和ATP合成的影响。结果显示，BF2-A/B都能结合基因组DNA，BF2-B的结合能力比BF2-A强，并且都趋向特异性结合基因组DNA上的2个区段，其中1个是23S rRNA基因上的1段保守序列，衍生肽还影响细胞的氧化磷酸化过程，显著抑制细胞呼吸作用和ATP合成， BF2-B的抑制比BF2-A更强烈。研究结果说明BF2-A/B是通过渗透细胞膜进入细胞质，特异性结合基因组DNA上的2个区段，并抑制细胞呼吸作用和ATP合成而杀菌。BF2-B结合DNA的能力更强，抑制细胞呼吸作用和ATP合成的能力更强，因此具有更强的抑菌活性。

Abstract:

In this paper，the bactericidal mechanism of BF2-A/B，two analogues of antimicrobial peptide BuforinⅡ，specific binding to Staphylococcus aureus genomic DNA had been researched. The results of atomic Force Microscope scan and gel retardation assay showed that both peptides could strongly bind to DNA，and the ability of BF2-B binding to DNA was stronger than that of BF2-A. The results of DNaseⅠ footprinting assay showed that both peptides tended to specifically bind to two domains of S.aureusgenomic DNA，and one of them probably was a fragment of consensus sequence located on the 23S rRNA gene. Furthermore，the results of interference assay to the oxidation phosphorylation of cells suggested that both peptides could obviously inhibit the respiration of S.aureus，inducing a drop in ATP content. Interestingly，BF2-B induced a more remarkable inhibition than BF2-A. The results of researches demonstrated that the powerful antimicrobial activities of both peptides were closely related to penetrate the cytoplasma membrane then specifically bind to two domains of genomic DNA，and inhibit the cell respiration and ATP synthesis. The increase of ability of binding to DNA and inhibiting cell respiration and ATP synthesis explained why BF2-B displayed more excellent antimicrobial activity for S.aureus.

中图分类号:

S852.42

郝刚，乐国伟，施用晖. 抗菌肽BuforinⅡ衍生物特异性结合金黄色葡萄球菌DNA抑菌机理的研究[J]. 畜牧兽医学报, 2014, 45(7): 1154-1161.

HAO Gang，LE Guo-wei，SHI Yong-hui. The Bactericidal Mechanism of Two Analogues of the Antimicrobial Peptide BuforinⅡSpecific Binding to Staphylococcus aureus Genomic DNA[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(7): 1154-1161.

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抗菌肽BuforinⅡ衍生物特异性结合金黄色葡萄球菌DNA抑菌机理的研究

The Bactericidal Mechanism of Two Analogues of the Antimicrobial Peptide BuforinⅡSpecific Binding to Staphylococcus aureus Genomic DNA

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