丙氨酸扫描法探究Cn-AMP2肽结构与抗菌活性关系  

Relationship between structure and antimicrobial activity of the Cn-AMP2 peptide by alanine scanning method

作　　者：李文勇 齐海燕[2,3] 衣同辉[4] 韩涵 沈焕泉 潘洪明[5] LI Wenyong;QI Haiyan;YI Tonghui;HAN Han;SHEN Huanquan;PAN Hongming(School of Medical Technology,Qiqihar Medical University,Qiqihar 161006,China;School of Chemistry and Chemical Engineering,Qiqihar University,Qiqihar 161006,China;Heilongjiang Industrial Hemp Processing Technology Innovation Center,Qiqihar University,Qiqihar 161006,China;Center for Health Inspection,Qiqihar Medical University,Qiqihar 161006,China;School of Basic Medicine,Qiqihar Medical University,Qiqihar 161006,China)

机构地区：[1]齐齐哈尔医学院医学技术学院,黑龙江齐齐哈尔161006 [2]齐齐哈尔大学化学与化学工程学院,黑龙江齐齐哈尔161006 [3]齐齐哈尔大学黑龙江省工业大麻加工技术创新中心,黑龙江齐齐哈尔161006 [4]齐齐哈尔医学院卫生检验中心,黑龙江齐齐哈尔161006 [5]齐齐哈尔医学院基础医学院,黑龙江齐齐哈尔161006

出　　处：《高师理科学刊》2023年第3期47-51,共5页Journal of Science of Teachers＇College and University

基　　金：国家自然科学基金项目(81641134);黑龙江省卫健委基金项目(20210202020365);黑龙江省大学生创新创业训练项目(202211230003)。

摘　　要：为研究抗菌肽Cn-AMP2的结构与抗菌活性之间的关系,采用丙氨酸扫描法进行突变,设计一系列抗菌肽Cn-AMP2丙氨酸扫描突变体.应用生物信息学法研究突变体的理化性质参数,测定抗菌肽Cn-AMP2和突变体肽对8种革兰阴性菌和革兰阳性菌的抗菌活性,并以此为基础对抗菌肽Cn-AMP2的构效关系进行深入研究.结果表明,丙氨酸突变后,分子量和净电荷数量无明显变化,但疏水力矩有明显变化.Cn-AMP2和突变体肽抗革兰阴性菌最低抑菌浓度(MIC)为16~354μmol/L,抗革兰阳性菌最低抑菌浓度为26~250μmol/L.圆二色光谱测定表明,Cn-AMP2和突变体肽在222 nm处的摩尔椭圆率分别为-1362,-401 deg·cm^(2)·dmol^(-1).NPN荧光探针实验表明,Cn-AMP2和突变体肽对细菌细胞外膜破坏强度有明显差异.实验结果表明,第3位丝氨酸、第4位酪氨酸、第8位丝氨酸和第11位甲硫氨酸为关键位点.In order to study the relationship between the structure and antimicrobial activity of antibacterial peptide Cn-AMP2,a series of alanine scanning mutants of antibacterial peptide Cn-AMP2 were designed by using alanine scanning method.The physical and chemical properties of the mutants were studied by the bioinformatic methods.The antimicrobial activities of antibacterial peptide Cn-AMP2 and mutant peptide against 8 kinds of Gram-negative and Gram-positive bacteria were determined,and the structure-activity relationship of antibacterial peptide Cn-AMP2 was further studied on this basis.The experimental results showed that the molecular weight and the number of net charges did not change significantly after alanine mutation,but the hydrophobic moment changed significantly.The MIC of Cn-AMP2 and mutant peptide against Gram-negative bacteria is 16~354μmol/L,and against Gram-positive bacteria is 26~250μmol/L.Circular dichroism spectroscopy showed that the molar ellipticity of Cn-AMP2 and mutant peptide at 222 nm was-1362,-401 deg·cm^(2)·dmol^(-1).NPN fluorescence probe experiment showed that Cn-AMP2 and mutant peptide had significant differences in the destruction intensity of bacterial outer membrane.The results showed that serine 3,tyrosine 4,serine 8 and methionine 11 were the key sites.

关 键 词：抗菌肽 丙氨酸扫描法 Cn-AMP2 关键位点 

分 类 号：Q5[生物学—生物化学]