Individual N-glycans added at intervals along the stalk of the Nipah virus G protein prevent fusion but do not block the interaction with the homologous F protein.
Individual N-glycans added at intervals along the stalk of the Nipah virus G protein prevent fusion but do not block the interaction with the homologous F protein.
复制标题
沿着尼帕病毒 G 蛋白的茎部间隔添加的单个 N-聚糖可防止融合，但不会阻止与同源 F 蛋白的相互作用。
DOI：
10.1128/jvi.03084-12
复制
发表时间：
2013
影响因子：
5.4
通讯作者：
Iorio,RonaldM
中科院分区：
文献类型：
--
作者：
Zhu,Qiyun;Biering,ScottB;Mirza,AnneM;Grasseschi,BrittanyA;Mahon,PaulJ;Lee,Benhur;Aguilar,HectorC;Iorio,RonaldM
大多数副粘病毒促进膜融合需要病毒附着和融合(F)蛋白之间的相互作用，以使前者能够与受体结合，从而触发后者的激活以进行融合。大量研究表明，新城疫病毒(NDV)、血凝素神经氨酸酶(HN)和麻疹病毒(MV)血凝素(H)蛋白上的F-相互作用位点完全位于这些蛋白的茎区。事实上，新城疫病毒HN和MV H的茎残基可能介导了F的相互作用。然而，尽管进行了广泛的研究，但尼帕病毒(Nipah Virus，NIV)G附着糖蛋白上的F-相互作用位点(S)仍未被鉴定。在这项研究中，我们在沿着NIV G蛋白的茎上间隔开的几个位置引入了单独的N连接糖基化位点。7个引入的位点中有5个被利用，这是由于电泳迁移率的延迟而建立的。尽管表面表达、ewitinB2结合和寡聚作用与野生型蛋白相似，但添加的五个N-糖链中有四个完全消除了G蛋白在促进融合方面补充同源F蛋白的能力。膜最近端添加的N-聚糖会减少80%的融合。然而，与类似的新城疫病毒HN和MV H突变体不同，NIV G糖基化茎突变体保留了与F结合的能力，表明这些突变体的融合缺陷不是由于阻止G-F相互作用造成的。这些发现表明，G-F相互作用并不完全由G的茎结构域介导，而可能比HN/H-F的作用更复杂。
The promotion of membrane fusion by most paramyxoviruses requires an interaction between the viral attachment and fusion (F) proteins to enable receptor binding by the former to trigger the activation of the latter for fusion. Numerous studies demonstrate that the F-interactive sites on the Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) and measles virus (MV) hemagglutinin (H) proteins reside entirely within the stalk regions of those proteins. Indeed, stalk residues of NDV HN and MV H that likely mediate the F interaction have been identified. However, despite extensive efforts, the F-interactive site(s) on the Nipah virus (NiV) G attachment glycoprotein has not been identified. In this study, we have introduced individual N-linked glycosylation sites at several positions spaced at intervals along the stalk of the NiV G protein. Five of the seven introduced sites are utilized as established by a retardation of electrophoretic mobility. Despite surface expression, ephrinB2 binding, and oligomerization comparable to those of the wild-type protein, four of the five added N-glycans completely eliminate the ability of the G protein to complement the homologous F protein in the promotion of fusion. The most membrane-proximal added N-glycan reduces fusion by 80%. However, unlike similar NDV HN and MV H mutants, the NiV G glycosylation stalk mutants retain the ability to bind F, indicating that the fusion deficiency of these mutants is not due to prevention of the G-F interaction. These findings suggest that the G-F interaction is not mediated entirely by the stalk domain of G and may be more complex than that of HN/H-F.