Payne L
J Virol. 1978 Jul;27(1):28-37. doi: 10.1128/JVI.27.1.28-37.1978.
Extracellular enveloped vaccinia (EEV) virus grown in SIRC and in HeLa cells was purified by consecutive equilibrium centrifugations in sucrose and cesium chloride gradients. A higher degree of purity was obtained with virus material prepared in SIRC cells. The polypeptides of purified EEV and INV (intracellular naked vaccinia) virus were compared in polyacrylamide slab gel electrophoresis. Three proteins (200,000 molecular weight [200K], 95K, and 13K) detected in HeLa-derived INV were absent in EEV. In addition, two INV proteins (65K and 30K) occurred in reduced concentrations in EEV, white another INV protein (27K) was increased in EEV. INV from SIRC cells showed similar alterations of these proteins (with the exception of the 30K and 13K proteins). Detergent treatment, ether extraction, and Pronase treatment showed that these six proteins are located at the surface of INV and are not cecessary for infectivity. Eight proteins (210K, 110K, 89K, 42K, 37K, 21.5K, 21K, and 20K) were detected in EEV that were absent from inv. Brij-58 treatment was employed to remove the envelope from EEV, resulting in the formation of naked particles and an envelope fraction which were separated on cesium chloride gradients. The envelope fractions contained all eight proteins. Seven of the eight proteins were glycoproteins, with the 37K protein being the only unglycosylated protein. It is concluded that a processing of surface INV particle proteins occurs during evelopment. The resultant EEV particle is comprised of an INV particle with a modified surface composition enclosed in an envelope containing virus-specific proteins unique to EEV.
在SIRC细胞和HeLa细胞中培养的细胞外被膜痘苗病毒（EEV），通过在蔗糖和氯化铯梯度中连续平衡离心进行纯化。用SIRC细胞制备的病毒材料获得了更高的纯度。在聚丙烯酰胺平板凝胶电泳中比较了纯化的EEV和INV（细胞内裸露痘苗病毒）病毒的多肽。在HeLa来源的INV中检测到的三种蛋白质（分子量200,000 [200K]、95K和13K）在EEV中不存在。此外，两种INV蛋白（65K和30K）在EEV中的浓度降低，而另一种INV蛋白（27K）在EEV中增加。来自SIRC细胞的INV显示出这些蛋白质的类似变化（30K和13K蛋白质除外）。去污剂处理、乙醚提取和链霉蛋白酶处理表明，这六种蛋白质位于INV表面，对感染性不是必需的。在EEV中检测到八种蛋白质（210K、110K、89K、42K、37K、21.5K、21K和20K），而在INV中不存在。采用Brij - 58处理从EEV中去除包膜，导致形成裸露颗粒和包膜组分，它们在氯化铯梯度上分离。包膜组分包含所有八种蛋白质。这八种蛋白质中的七种是糖蛋白，37K蛋白是唯一未糖基化的蛋白质。得出的结论是，在发育过程中发生了表面INV颗粒蛋白的加工。所得的EEV颗粒由表面组成经过修饰的INV颗粒组成，该颗粒被包裹在一个含有EEV特有的病毒特异性蛋白的包膜中。
