不同生物I型磷脂酶A2N末端衍生肽抗菌作用的研究.docxVIP

不同生物I型磷脂酶A2N?末端衍生肽抗菌作用的研究 陶廷婷史晓雄梁宁牛(通讯作者) (广西医科大学附属肿瘤医院药学部广西南宁530021) 【摘要】目的：研究人、猩猩、猪、小鼠、大鼠五种生物I型磷脂酶A2 (phospholipase A2, PLA2) N末端衍生肽在体外的杀菌效应,了解它们的结构与 功能的关系。方法:根据人、猩猩、猪、小鼠、大鼠五种生物I型PLA2N-端第1-15 个氨基酸残基为模板，分别合成衍生多肽AVW、ALW、AVN (因人与黑猩猩、大 鼠与小鼠I型PLA2N-端第1-15个氨基酸残基序列相同，最终合成3条衍生多肽)。 釆用琼脂铺板计数法，将不同浓度的多肽分别与革兰阳性菌金黄色葡萄球菌和革 兰阴性菌大肠杆菌在37°C孵育2.5h,然后铺板并置于37°C恒温箱培养18?24h, 记录每一块琼脂板上的菌落形成数，并计算多肽杀菌率。结果：衍生多肽AVW、 ALW、AVN对革兰阳性菌金黄色葡萄球菌和革兰阴性菌大肠杆菌均有杀菌活性, AVW和AVN对金黄色葡萄球菌和大肠杆菌杀菌作用较强，ALW对G+菌及G■菌 杀菌活性较弱。结论:不同生物I型PLA2N-末端衍生肽对G+菌和G?菌均有杀菌活 性，而具有较多碱性氨基酸衍牛肽杀菌活性较强。 【关键词】I型PLA2 N■端衍牛肽杀菌活性构效关系 【中图分类号】R919 【文献标识码】A 【文章编号】2095-1752 (2014) 12-0051-02 A Study on Bactericidal Activity of Group I Phospholipase A2 N-terminal Derived Peptides from Different Animals TAO Ting-ting,SHI Xiao-xiong,LIANG Ning-sheng (Department of Pharmacy, The Affiliated Tumor Hospital of Guangxi Medical University, Nanning ,530021) 【Abstract] Object:To analysis and compare the bactericidal activity of the polypeptides derives from the N-terminal sequenee of group I phospholipase A2 in Homo sapiens、Pan troglodytes^ Sus scrofa Rattus norvegicus and Mus musculus in vitro, to un dersta nd the relati on ship betwee n the bactericidal activity of the derived peptide and molecular structure? Methods:According to the N-terminal 15 amino acid sequence of group I PLA2 on the five mammal, The AVW, ALW and AVN were synthesized (because of the sequenee of group I PLA2 from the Homo sapiens and the Pan troglodytes, the Rattus norvegicus and the Mus musculus were the same ,so only 3 polypeptides were synthesized). Different concentrations of peptide incubated with Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli) in a water bath at 37°C for 2.5 h. The reaction solution mixed with agar solution and poured plates. The plates were incubated at a temperature of 37°C for 18-24 h, then were took out to count the colony-forming units (CFU) of each agar plate. The bactericidal rate was calculated .Results:3 pepti