腮腺炎病毒F蛋白七肽重复序列的表达、纯化及特性分析  

Expression and Purification of Heptad Repeat Region of the Mumps Virus F Protein and Analysis of Characteristics

作　　者：刘月永[1] 冯明光[1] 朱杰青[2] 姜立杰[3] 田波[2] 

机构地区：[1]浙江大学生命科学学院微生物研究所,杭州310029 [2]中国科学院微生物研究所分子病毒室,北京100080 [3]北京农林科学院林果所,北京100093

出　　处：《生物工程学报》2004年第3期377-381,共5页Chinese Journal of Biotechnology

基　　金：国家"973"计划资金资助 (No .G19990 75 60 2 )~~

摘　　要：副粘病毒F蛋白的两段七肽重复序列 (HR1和HR2 )在病毒侵染细胞的过程中相互作用形成热稳定的富含α螺旋的异源二聚体 ,此结构的形成引起病毒囊膜与细胞膜的并置而最终导致膜融合的发生。腮腺炎病毒(Mumpsvirus ,MuV)属于副粘病毒科 ,腮腺炎病毒属 ,可能利用与其他副粘病毒相似的侵染机制。本研究对MuV融合蛋白的HR区进行了计算机程序预测 ,并利用大肠杆菌GST融合表达系统对MuVF蛋白HR1和HR2两段多肽进行了表达和纯化 ,通过GSTpull_down实验证实HR1和HR2多肽在体外能够相互作用 ,凝胶过滤层析证明HR1、HR2多肽能够形成多聚体 ,说明MuVF蛋白的HR区的相互作用可能是其发挥融合功能的关键因素。Two Heptad repeat motifs (HR1 and HR2) from paramyxoviruses F protein could form thermostable heterodimers containing high α-helix while virus infected host cell. Following that the viral membrane and the host cell membrane were juxtaposed, which leads to membrane fusion. Mumps virus (MuV) is a member of the genus Rubulavirus in the family of Paramyxoviridae. MuV could use similar infection mechanism as well as other paramyxoviruses. In this study the HR1 and HR2 regions of MuV F protein were predicted by a computer program and expressed in E.coli with the GST fusion expression system. The GST fusion or GST-removed proteins were purified with Gluthathion Sepharose 4B Column. GST pull-down experiment suggested the interaction of HR1and HR2 peptides, and analysis of gel filtration showed two peptides could form multimer, which indicates that the HR regions of MuV F protein may play an important role in virus fusion.

关 键 词：腮腺炎病毒(Mumps virus MuV) F蛋白 七肽重复序列(HR) GST pull-down 

分 类 号：R346[医药卫生—基础医学]