Six-helix bundle assembly and characterization of heptad repeat regions from the F protein of Newcastle disease virus
Six-helix bundle assembly and characterization of heptad repeat regions from the F protein of Newcastle disease virus
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DOI：
10.1099/0022-1317-83-3-623
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发表时间：
2002-03-01
影响因子：
3.8
通讯作者：
Gao, GF
中科院分区：
文献类型：
--
作者：
Yu, M;Wang, EX;Gao, GF
副粘病毒可能采用与其他包膜病毒相似的融合机制，其中在包膜融合蛋白的七肽重复（HR）区域中融合后形成反平行的六螺旋束结构。为了解副粘病毒科纽卡斯尔病病毒（NDV）的融合机制并筛选其融合抑制剂，我们构建了一个E.将F蛋白HR 1和HR 2区分别表达为GST融合蛋白的大肠杆菌系统。纯化的裂解HR 1和HR 2随后组装成一个稳定的六螺旋束异源三聚体复合物。此外，GST融合蛋白和裂解的HR 2都显示出病毒-细胞融合抑制活性（IC 50为1.07-2.93 μ M）。GST-HR 2融合蛋白的溶解度远高于相应肽的溶解度。因此，这为大规模生产作为病毒融合抑制剂的HR肽提供了合理的方法。
Paramyxoviruses may adopt a similar fusion mechanism to other enveloped viruses, in which an antiparallel six-helix bundle structure is formed post-fusion in the heptad repeat (HR) regions of the envelope fusion protein. In order to understand the fusion mechanism and identify fusion inhibitors of Newcastle disease virus (NDV), a member of the Paramyxoviridae family, we have developed an E. coli system that separately expresses the F protein HR1 and HR2 regions as GST fusion proteins. The purified cleaved HR1 and HR2 have subsequently been assembled into a stable six-helix bundle heterotrimer complex. Furthermore, both the GST fusion protein and the cleaved HR2 show virus-cell fusion inhibition activity (IC50 of 1.07-2.93 muM). The solubility of the GST-HR2 fusion protein is much higher than that of the corresponding peptide. Hence this provides a plausible method for large-scale production of HR peptides as virus fusion inhibitors.