ISOLATION OF CORONAVIRUS ENVELOPE GLYCOPROTEINS AND INTERACTION WITH THE VIRAL NUCLEOCAPSID
ISOLATION OF CORONAVIRUS ENVELOPE GLYCOPROTEINS AND INTERACTION WITH THE VIRAL NUCLEOCAPSID
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DOI：
10.1128/jvi.33.1.449-462.1980
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发表时间：
1980-01-01
影响因子：
5.4
通讯作者：
BEHNKE, J
中科院分区：
文献类型：
--
作者：
STURMAN, LS;HOLMES, KV;BEHNKE, J
通过在 4℃下用 Nonidet P-40 溶解病毒膜来分离冠状病毒 A59 的 2 个包膜糖蛋白和病毒核衣壳。 C随后进行蔗糖密度梯度沉降。分离的 E2 由 pplomer 的玫瑰花结组成，而 E1（膜糖蛋白）是不规则且无定形的。在某些条件下，Nonidet P-40 破坏的病毒颗粒的成分之间会发生显着的相互作用。 Nonidet P-40 破坏的病毒在 37°C 下孵育。 C导致病毒糖蛋白之一E1和病毒核衣壳之间形成复合物。这是由 E1 的温度依赖性构象变化引起的，导致 E1 聚集并与核衣壳中的病毒 RNA 相互作用。 E1也结合rRNA。 E1-核衣壳复合物可以在蔗糖和 Renografin 密度梯度上与天然病毒核衣壳区分开来。将膜糖蛋白 E1 与膜糖蛋白 E2 分离，可以制备针对这些分离蛋白的抗血清。提出了一个模型来描述冠状病毒 A59 病毒粒子中 3 个主要结构蛋白与病毒包膜和 RNA 的关系。
The 2 envelope glycoproteins and the viral nucleocapsid of the coronavirus A59 were isolated by solubilization of the viral membrane with Nonidet P-40 at 4.degree. C followed by sucrose density gradient sedimentation. Isolated E2 consisted of rosettes of peplomers, whereas E1, the membrane glycoprotein, was irregular and amorphous. Under certain conditions significant interactions occurred between components of Nonidet P-40-disrupted virions. Incubation of the Nonidet P-40-disrupted virus at 37.degree. C resulted in formation of a complex between one of the viral glycoproteins, E1, and the viral nucleocapsid. This was caused by a temperature-dependent conformational change in E1, resulting in aggregation of E1 and interaction with the viral RNA in the nucleocapsid. E1 also bound rRNA. The E1-nucleocapsid complexes can be distinguished on sucrose and Renografin density gradients from native viral nucleocapsids. The separation of the membrane glycoprotein E1 from the peplomeric glycoprotein E2 permitted preparation of antisera against these isolated proteins. A model is proposed for the arrangement of the 3 major structural proteins in the coronavirus A59 virion in relation to the viral envelope and RNA.