麻疹病毒包膜糖蛋白与受体的相互作用及其相互作用功能位点研究

发布时间：2018-01-30 11:13

本文关键词： 麻疹病毒血凝素蛋白融和蛋白受体 SLAM 免疫共沉淀 pull-down激光共聚焦 co-capping 流式细胞计数术噬菌体展示系统　出处：《武汉大学》2005年硕士论文　论文类型：学位论文

【摘要】：麻疹病毒血凝素蛋白(Hemagglutinin protein,H)是介导麻疹病毒吸附、侵入宿主细胞的关键分子。CD46是被鉴定的麻疹病毒的第一个受体。人细胞表面的SLAM(signaling lymphocytic activation molecule)分子是2000年鉴定的麻疹病毒的第二受体。2001年,本室李凌云博士从绒猴B95a细胞cDNA文库中发现了与SLAM高度同源的蛋白BIP,并且验证了它也能作为麻疹病毒的受体,我们称为mSLAM。为了进一步研究麻疹病毒第二受体SLAM(或mSLAM)在麻疹病毒侵染宿主过程中的作用,我们着重分析了麻疹病毒两种糖蛋白H蛋白,F蛋白和SLAM的相互作用。在本文研究结果和前人研究的基础上,为阐明麻疹病毒的侵染机制,提出了麻疹病毒H蛋白和F蛋白与SLAM受体之间形成三聚体模型。首先我们利用免疫共沉淀方法在体内证实了麻疹病毒的两种糖蛋白H和F与细胞表面受体SLAM之间的相互作用。然后利用pull-down进一步证实了三种蛋白在体外的相互结合,确定了三种蛋白之间相互作用并形成三聚体的蛋白复合物。通过转染HeLa细胞在细胞形态学上对麻疹病毒的血凝集素蛋白H和融合蛋白F的功能进行了研究。发现麻疹病毒的侵染融合必须在三种蛋白的共同作用下才能顺利进行。同时,利用激光共聚焦的方法,发现了这两种糖蛋白在SLAM的介导下,在细胞膜上的分布发生变化形成co-cap现象。从而确认了三者在细胞内部通过其共同形成的三聚体发挥功能。
[Abstract]:Measles virus hemagglutinin protein (HH) mediates measles virus adsorption. CD46, the key molecule that invades host cells, is the first receptor of the identified measles virus. Signaling lymphocytic activation molecule is the second receptor of measles virus identified in 2000. 2001. Dr Li Lingyun identified a highly homologous SLAM protein from the rhesus monkey B95a cell cDNA library and confirmed that it can also act as a receptor for measles virus. We call mSLAM.To further study the role of SLAM (or mSLAM), the second receptor of measles virus, in the process of measles virus infecting the host. The interaction between two glycoprotein H protein F protein and SLAM of measles virus was analyzed. Based on the results of this study and previous studies, the infection mechanism of measles virus was elucidated. A trimer model between H protein and F protein of measles virus and SLAM receptor was proposed. The interaction between the two glycoproteins H and F of measles virus and the cell surface receptor SLAM was confirmed by immunoprecipitation in vivo. Then, the interaction between H and F of measles virus and the cell surface receptor SLAM was further confirmed by pull-down. The interaction of three proteins in vitro. The interaction of three proteins and the formation of trimer protein complex were identified. The functions of hemagglutinin protein H and fusion protein F of measles virus were studied by transfection of HeLa cells in cell morphology. It was found that the infection and fusion of measles virus must be carried out successfully under the combined action of three proteins. Using laser confocal method, we found that these two glycoproteins were mediated by SLAM. The distribution on the cell membrane changed to form the co-cap phenomenon, which confirmed the function of the three trimers in the cell.
【学位授予单位】：武汉大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R373

【共引文献】