Shotgun质谱筛选与猪瘟病毒囊膜蛋白E^rns相互作用的候选宿主蛋白  

Shotgun screening of cellular proteins potentially interacting with E^(rns) of classical swine fever virus

作　　者：尹凝[1,2] 蒋大良[3] 龚文杰[2] 郭焕成[2] 谢金鑫[2] 夏乐乐[1,2] 余兴龙[3] 涂长春[2] 

机构地区：[1]吉林农业大学动物科技学院,吉林长春130118 [2]军事医学科学院军事兽医研究所,吉林长春130122 [3]湖南农业大学动物医学院,湖南长沙410128

出　　处：《中国预防兽医学报》2013年第10期799-803,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金重点项目(31130052)

摘　　要：猪瘟病毒(CSFV)Erns蛋白是瘟病毒属成员特有的囊膜糖蛋白,在病毒吸附和侵入靶细胞过程中发挥重要作用。本研究利用Erns特异性抗体对CSFV石门强毒株感染的PK-15细胞进行免疫共沉淀(Co-IP),对获得的蛋白复合物进行Shotgun质谱鉴定。结果显示,与阴性对照相比,病毒感染细胞样品中含有199种差异蛋白,其中可能包含与Erns作用的宿主细胞蛋白。同时,实验还对Co-IP蛋白样品进行SDS-PAGE,切取差异条带进行质谱分析以进一步验证Shotgun技术的蛋白筛选结果。通过Co-IP和Shotgun质谱分析,本研究筛选出一些可能在CSFV生命周期中与Erns互作用的宿主蛋白,为进一步研究CSFV在宿主细胞中的感染和复制的分子机制奠定基础。Envelope glycoprotein E-ms of classical swine fever virus （CSFV）, a unique envelope protein of the pestivirus members, plays an important role in the process of virus adsorption and entry into target cells. To screen Er-ms interacting with host proteins, specific antibody against Er-ms was used for co-immunoprecipitation （Co-IP） to isolate E-ms interacting cellular proteins in CSFV-infected PK-15 cells, the obtained proteins were then identified by Shotgun. Results showed that 199 differential host proteins were identified in CSFV-infected sample compared to the negative control, some of which might be the interacting proteins of E-ms. To further confirm the protein screening results by Shotgun, the protein complex obtained by Co-IP were subjected to SDS-PAGE and the differential band was analyzed by mass spectrometry. Overall, this study is the first time to study the interaction between CSFV E~ and cellular proteins by Co-IP and Shotgun, and the screened proteins might play a pivotal role in the life cycle of CSFV. These results might provide a basis for further unraveling the molecular mechanism of CSFV pathogenesis.

关 键 词：猪瘟病毒 蛋白质相互作用 SHOTGUN Erns 宿主细胞蛋白 

分 类 号：S852.65[农业科学—基础兽医学]