A 45,000-M(r) glycoprotein in the Sendai virus envelope triggers virus-cell fusion
A 45,000-M(r) glycoprotein in the Sendai virus envelope triggers virus-cell fusion
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仙台病毒包膜中的 45,000 M(r) 糖蛋白触发病毒与细胞融合
DOI：
10.1128/jvi.71.9.6398-6406.1997
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发表时间：
1997
影响因子：
5.4
通讯作者：
D. P. Sarkar
中科院分区：
文献类型：
--
作者：
M. Kumar;M. Hassan;S. Tyagi;D. P. Sarkar
仙台病毒包膜缺乏血凝素-神经氨酸酶，但含有融合蛋白（F-病毒体）的制备。F-病毒体表现出明显的丝氨酸蛋白酶活性，在中性pH值。电泳分析的F-病毒体的蛋白质分布在非还原条件下，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和等电聚焦，导致识别一个以前未知的糖蛋白与45,000（45 K蛋白）的相对分子量与F蛋白。45 K蛋白的身份，从F蛋白不同，建立了用F和45 K特异性抗体的蛋白质印迹分析。这种45 K蛋白通过非共价疏水相互作用与F蛋白形成连接，如其对尿素处理的敏感性所证明的，并且它对于F病毒体的蛋白水解活性以及对于病毒包膜与宿主细胞膜的融合是必不可少的。该蛋白的N-末端序列分析（前11个氨基酸）显示与黄病毒NS 3丝氨酸蛋白酶具有很强的同源性（> 90%），但与任何仙台病毒蛋白都没有相似性。根据N-末端序列，设计对应于正义和反义DNA序列的寡核苷酸。用这些寡核苷酸与病毒和宿主基因组的斑点杂交和引物延伸证实了该蛋白的宿主来源。此外，有限的蛋白水解酶消化的目标膜，导致与它的病毒融合显着抑制。在这些结果的基础上，我们假设的F蛋白诱导的膜融合的分子机制的模型，这可能提供了一个理论基础，为其他副粘病毒。
Sendai virus envelopes devoid of hemagglutinin-neuraminidase but containing the fusion protein (F-virosomes) were prepared. F-virosomes exhibited discernible serine protease activity at neutral pH. Electrophoretic analysis of the protein profile of the F-virosomes under nonreducing conditions, by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing, led to the identification of a previously unknown glycoprotein with a relative molecular weight of 45,000 (45K protein) associated with the F protein. The identity of the 45K protein, as distinct from F protein, was established by Western blot analysis with F- and 45K-specific antibodies. This 45K protein forms a nexus with the F protein through noncovalent hydrophobic interactions, as proved by its sensitivity to urea treatment, and it is essential for the proteolytic activity of the F-virosomes as well as for the fusion of the viral envelope with host cell membrane. N-terminal sequence analysis (first 11 amino acids) of this protein showed strong homology (> 90%) to flavivirus NS3 serine proteases but no similarity to any of the Sendai viral proteins. On the basis of the N-terminal sequence, oligonucleotides were designed corresponding to the sense and antisense DNA sequences. Dot blot hybridization and primer extension with these oligonucleotides with the viral and the host genome confirmed the host origin of this protein. Further, the limited proteolytic digestion of the target membrane resulted in significant inhibition of viral fusion with it. On the basis of these results, we postulate a model for the molecular mechanism of F protein-induced membrane fusion, which may provide a rationale for other paramyxoviruses.
病毒包膜融合后仙台病毒蛋白与红细胞膜细胞质表面的相互作用。
DOI：
--
发表时间：
1981
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Caldwell,SE;Lyles,DS
通讯作者：
Lyles,DS