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Studies on protein regulation of virus-mediated membrance fusion

病毒介导的膜融合蛋白调控研究

基本信息

批准号：
05680608
负责人：
TSURUDOME Masato
金额：
$ 1.28万
依托单位：
Mie University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

1.Previosly, we had obtained four escape mutants under pressure of an anti-HN monoclonal antibody (MAb), which inhibited cell fusion induced by human parainfluenza type 2 virus (HPIV2). In the present study, further analyzes have proved that the MAb recognizes 83Asn and 91Lys on the HN molecule.2.The HN and F proteins of three paramyxoviruses (HPIV2, HPIV4, and SV41) were expressed in cells by transfecting them with recombinant plasmid encoding each protein gene. Coexpression of HPIV2 HN with HPIV2 F protein resulted in induction of prominent cell fusion. HPIV2 HN could also promote cell fusion when coexpressed with SV41 F or HPIV4 F proteins, although at lower extents. SV41 HN or HPIV4 HN did not induce cell fusion by coexpressing with HPIV2 F protein.3.Based on the above observation, futher analyzes were performed by constructing and expressing 32 chimeric proteins of HPIV2 HN and SV41 HN proteins. It was revealed that two regions on the HPIV2 HN protein ranging from 37Ser to 91Ser (region A) and ranging from 148Gly to 20Ala (region B) are involved in functional molecular interaction with the F protein, which is a prerequiste for cell fusion induction. On the other hand, similar analyzes of chimeric F proteins have indicated that two regions on HPIV2 F protein ranging from 128Ala to 227Ala (region Y) and from 430Tur to 551Ser (region Z) are involved in the functional interaction with the HN protein.4.These results suggest that molecular interaction between HN and F proteins are indispensable for cell fusion, and each protein has two regions which are involed in the interaction. It is notewothy that one of the two regions of each protein includes the transmembrane domains which may be in close proximity to the fusion site.

1.在此之前，我们在抗hn单克隆抗体（MAb）的压力下获得了四个逃逸突变体，它们可以抑制人副流感2型病毒（HPIV2）诱导的细胞融合。在本研究中，进一步的分析证明了MAb可以识别HN分子上的83Asn和91Lys。用编码蛋白基因的重组质粒转染3种副粘病毒（HPIV2、HPIV4和SV41），在细胞中表达了HN蛋白和F蛋白。HPIV2 HN与HPIV2 F蛋白的共表达诱导了显著的细胞融合。当与SV41 F或HPIV4 F蛋白共表达时，HPIV2 HN也能促进细胞融合，尽管程度较低。2 . sv41hn或HPIV4 HN与HPIV2 F蛋白共表达不诱导细胞融合。在此基础上，构建并表达32个HPIV2 HN和SV41 HN蛋白嵌合蛋白进行进一步分析。结果表明，HPIV2 HN蛋白上37Ser ~ 91Ser （A区）和148Gly ~ 20Ala （B区）两个区域参与了与F蛋白的功能性分子相互作用，这是诱导细胞融合的先决条件。另一方面，对嵌合F蛋白的类似分析表明，HPIV2 F蛋白上的两个区域，从128Ala到227Ala （Y区）和从430Tur到551Ser （Z区）参与了与HN蛋白的功能相互作用。这些结果表明，HN蛋白和F蛋白之间的分子相互作用对于细胞融合是必不可少的，并且每个蛋白都有两个参与相互作用的区域。值得注意的是，每个蛋白质的两个区域之一包括跨膜结构域，它可能靠近融合位点。