LIPID COMPOSITION OF PURIFIED VESICULAR STOMATITIS VIRUSES
LIPID COMPOSITION OF PURIFIED VESICULAR STOMATITIS VIRUSES
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DOI：
10.1128/jvi.7.1.59-70.1971
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发表时间：
1971-01-01
影响因子：
5.4
通讯作者：
WAGNER, RR
中科院分区：
文献类型：
--
作者：
MCSHARRY, JJ;WAGNER, RR
本文描述了生产水泡性口炎（VS）病毒的方法，其纯度足以进行可靠的化学分析。从感染细胞中释放的VS病毒粒子通过聚乙二醇沉淀、柱层析、速率区离心和平衡离心的顺序步骤浓缩和纯化至少150倍。在l细胞中繁殖的Indiana血清型（vsind病毒）含有3%的核糖核酸，64%的蛋白质，13%的碳水化合物和20%的脂质；胆固醇与磷脂的摩尔比为0.6或更高。薄层色谱未发现异常的中性脂或磷脂，气液色谱未发现异常的脂肪酸掺入VS病毒粒子。在l细胞中生长的具有抗原性的新泽西血清型病毒（VSNJvirus）除了中性脂质比例高于同样在l细胞中生长的VSNJvirus外，其脂质谱与在l细胞中生长的VSNJvirus相似。当比较在鸡胚细胞中生长的vsind和vsnj病毒的脂质组成时，这种差异不那么明显，但在任何一种细胞类型中生长的vsnj病毒总是比vsind病毒含有更多的中性脂质，而不是胆固醇。在l细胞或鸡胚细胞中生长的vsind和vsnjv病毒的脂质组成更接近于质膜，而不是整个细胞。一个一致的发现是，与未感染的全l细胞和鸡胚细胞或其质膜相比，两种VS病毒中磷脂酰乙醇胺和鞘磷脂含量相对较高，磷脂酰胆碱含量相对较低。现有的质膜分离方法不足以对VS病毒粒子与其宿主细胞的质膜脂质进行结论性比较。然而，获得的数据与两个假设是一致的：(i) VS病毒的脂质组成主要反映了它们的膜成熟位点，（ii）插入细胞膜的新合成病毒蛋白影响了被选择并入病毒膜的磷脂和中性脂质的比例。
Methods are described for the production of vesicular stomatitis (VS) virus of sufficient purity for reliable chemical analysis. VS virions released from infected cells were concentrated and purified at least 150-fold by sequential steps of precipitation with polyethylene glycol, column chromatography, rate zonal centrifugation, and equilibrium centrifugation. The Indiana serotype (VSIndvirus) propagated in L-cells was found to contain 3% ribonucleic acid, 64% protein, 13% carbohydrate, and 20% lipid; the molar ratio of cholesterol to phospholipid was 0.6 or greater. Thin-layer chromatography revealed no unusual neutral lipids or phospholipids and gas-liquid chromatography revealed no unusual fatty acids incorporated into VS virions. The antigenically distinct New Jersey serotype (VSNJvirus) grown in L-cells showed a similar lipid profile except that the proportion of neutral lipids was larger than in VSIndvirus also grown in L-cells. This differences was less pronounced when the lipid composition of VSIndand VSNJviruses grown in chick embryo cells was compared, but VSNJvirus grown in either cell type always contained larger amounts of neutral lipids other than cholesterol than did VSIndvirus. The lipid composition of both VSIndand VSNJviruses grown in L-cells or chick embryo cells more closely resembled that of plasma membrane than of whole cells. A consistent finding was the relatively large amounts of phosphatidylethanolamine and sphingomyelin and the relatively small amounts of phosphatidylcholine in both VS viruses compared with uninfected whole L-cells and chick embryo cells or their plasma membranes. The methods available for isolation of plasma membranes were inadequate for conclusive comparison of the lipids of VS virions with the lipids of the plasma membranes of their host cells. Nevertheless, the data obtained are consistent with two hypotheses: (i) the lipid composition of VS viruses primarily reflects their membrane site of maturation, and (ii) the newly synthesized viral proteins inserted into cell membranes influence the proportions of phospholipids and neutral lipids selected for incorporation into the viral membrane.