马尔堡病毒GP蛋白C肽的抑制活性及作用机理研究  

Inhibitory Activity and Reaction Mechanism of C Peptide from Marburg Virus Glycoprotein

作　　者：陈文文 郑文铝 余硕 花晨 戴秋云 CHEN Wen-Wen;ZHENG Wen-Lyu;YU Shuo;HUA Chen;DAI Qiu-Yun(Beijing Institute of Biotechnology,Beijing 100071;School of Basic Medical Sciences,Fudan University,Shanghai 200433,China)

机构地区：[1]军事医学研究院生物工程研究所,北京100071 [2]复旦大学基础医学院,上海200433

出　　处：《生物技术通讯》2020年第1期44-50,共7页Letters in Biotechnology

基　　金：国家传染病防治重大专项(2016ZX10004001)。

摘　　要：目的:研究马尔堡病毒(MARV)包膜糖蛋白(GP)C肽的抗该病毒进入活性及初步作用机制。方法:选取MARV的GP2蛋白上CHR区域序列(611~637残基),合成该序列多肽(C肽)及突变体,利用pVAX1-MGP与pNL-4.3-Luc-R-E-质粒包装假病毒体系,测定各肽对马尔堡假病毒融合的抑制活性,利用圆二色谱、凝胶色谱分析C肽及其突变体对MARV的N肽的相互作用。结果:C肽对马尔堡假病毒融合抑制的IC50为32.67μmol/L,C肽突变体的IC50为9.61~33.47μmol/L,抑制活性高于C肽或与其相当;C肽与N肽形成双螺旋结构,在C肽的7元螺旋体的b、f、c、g位引入离子相互作用氨基酸可提高多肽本身的α螺旋结构含量,一些突变可提高复合物的α螺旋结构含量,但C肽与N肽混合后形成的复合物含量不高。结论:MARV的GP蛋白C肽与N肽相互作用较弱,C肽的抗病毒活性较弱,但改造后可提高活性。Objective:To study the inhibitory activity and reaction mechanism of C peptide from Marburg virus(MARV)glycoprotein(GP).Methods:The sequence(aa 611~637)of the CHR region in GP2 of MARV was se lected,and the corresponding peptide(C peptide)and its mutants were synthesized.The pVAX1-MGP and pNL-4.3-Luc-R-E-plasmids were used to construct the pseudovirus system,the inhibitory activity of peptides to cell fu sion of Marburg pseudovirus was determined.The circular dichroism and high pressure gel chromatography were used to determine the interaction of the C peptide and its mutants with the N peptide of MARV GP2.Results:The IC50 of C peptide against Marburg pseudovirus is 32.67μmol/L,the IC50 of C peptide mutants ranges from 9.61~33.47μmol/L,which is higher than or similar to that of C peptide.The C peptide and N peptide can form a double helix structure,the introduction of electrostatic interaction at the position of b,f,c,and g of the C pep tide heptad repeat increases theα-helix content of the polypeptide itself,some mutants increase theα-helix con tent of the complexes,but the content ofα-helix of complexes is not high.Conclusion:The interaction of C peptide with N peptide of MARV GP is low,C peptide exhibits weak antiviral activity,but its mutants have higher activity than the original C peptide.

关 键 词：马尔堡病毒 包膜糖蛋白(GP) C肽 N肽 抗病毒活性 

分 类 号：Q503[生物学—生物化学] R392[医药卫生—免疫学]