PH-DEPENDENT FUSION BETWEEN THE FLAVIVIRUS WEST NILE AND LIPOSOMAL MODEL MEMBRANES

PH-DEPENDENT FUSION BETWEEN THE FLAVIVIRUS WEST NILE AND LIPOSOMAL MODEL MEMBRANES
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DOI:
10.1099/0022-1317-67-1-157
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发表时间:
1986-01-01
影响因子:
3.8
通讯作者:
PORTERFIELD, JS
PORTERFIELD, JS
中科院分区:
医学3区
文献类型:
--
作者:
GOLLINS, SW;PORTERFIELD, JS

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纯化的[3H]尿苷标记的西尼罗病毒(WNV)颗粒和含有RNA酶的脂质体之间的融合,通过将病毒RNA降解为三氯乙酸可溶性物质来测定。发现病毒与含有磷脂酰胆碱、磷脂酰乙醇胺、鞘磷脂和胆固醇(摩尔比为1:1:1:1.5)的脂质体的融合依赖于pH,最大融合发生在pH 6.7及以下。在pH6.6时融合迅速,在37 ℃下2分钟内基本完成。C.此时,约50%的病毒RNA已被降解,增加脂质体浓度或允许融合的时间仅略微增加该百分比。融合是依赖于温度,几乎是完全非泄漏,并不依赖于二价阳离子的存在。发现脂质体的脂质组成影响融合的最佳pH和观察到的最大融合程度。电镜观察脂质体与病毒颗粒的融合反应。
Fusion between purified [3H]uridine-labelled West Nile virus (WNV) particles and liposomes containing RNase, was assayed by degradation of the viral RNA to trichloroacetic acid-soluble material. Fusion of virus with liposomes containing phosphatidylcholine, phosphatidylethanolamine, sphingomyelin and cholesterol (at a molar ratio of 1:1:1:1.5) was found to be dependent on pH with maximum fusion occurring at pH 6.7 and below. At pH 6.6 fusion was rapid and was essentially complete within 2 min at 37.degree. C. At this time, approximately 50% of the viral RNA had been degraded and increasing the concentration of liposomes or time allowed for fusion increased this percentage only slightly. Fusion was dependent on temperature, was almost totally non-leaky and was not dependent on the presence of divalent cations. The lipid composition of liposomes was found to influence both the pH optimum for fusion and the maximum degree of fusion observed. Electron microscopy was used to visualize the fusion reaction between liposomes and virus particles.