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通过荧光去极化研究披膜病毒膜的微粘度:包膜蛋白和宿主细胞的影响

Microviscosity of togavirus membranes studied by fluorescence depolarization: influence of envelope proteins and the host cell.

作者信息

Moore N F, Barenholz Y, Wagner R R

出版信息

J Virol. 1976 Jul;19(1):126-35. doi: 10.1128/JVI.19.1.126-135.1976.

Abstract

The microviscosities of the hydrophobic regions of the membranes of intact Semliki forest and Sindbis viruses grown on BHK-21 cells, of liposomes derived from the extracted viral lipids, and of protease-treated virions were measured by fluorescence depolorization using the fluorescence probe 1, 6-diphenyl-1,3,5-hexatriene. The intact virus membranes were found to have a higher microviscosity than did virus-derived liposomes, indicating the viral envelope proteins contribute to microviscosity. However, protease-treated virus, devoid of protruding spikes but with residual lipophilic peptide tails, was found to have a microviscosity more similar to that of the intact virus than to that of protein-free liposomes. Sindbis virus grown in BHK-21 cells at 37 C had a much higher microviscosity than did Sindbis virus grown on Aedes albopicuts cells at 22 C. Sindbis virus grwon in A. albopictus and BHK-21 cells also gave higher microviscosity values than did the intact host cells. These data indicate that both the virion proteins and the cellular lipids selected during viral growth and maturation contribute to the increased microviscosity of togavirus membranes.

摘要

使用荧光探针1,6 - 二苯基 - 1,3,5 - 己三烯通过荧光去极化法测量了在BHK - 21细胞上生长的完整Semliki森林病毒和辛德毕斯病毒、从提取的病毒脂质衍生的脂质体以及经蛋白酶处理的病毒粒子膜疏水区域的微粘度。结果发现,完整病毒膜的微粘度高于病毒衍生的脂质体,这表明病毒包膜蛋白对微粘度有贡献。然而,经蛋白酶处理的病毒,虽然没有突出的刺突但仍有残留的亲脂性肽尾,其微粘度与完整病毒的微粘度更相似,而与无蛋白脂质体的微粘度不同。在37℃下于BHK - 21细胞中生长的辛德毕斯病毒的微粘度比在22℃下于白纹伊蚊细胞中生长的辛德毕斯病毒高得多。在白纹伊蚊和BHK - 21细胞中生长的辛德毕斯病毒的微粘度值也高于完整宿主细胞。这些数据表明,病毒生长和成熟过程中选择的病毒粒子蛋白和细胞脂质都有助于增加披膜病毒膜的微粘度。

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