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仙台病毒与带负电荷脂质体的相互作用:病毒诱导的羧基荧光素负载小单层囊泡的裂解

The interaction of Sendai virus with negatively charged liposomes: virus-induced lysis of carboxyfluorescein-loaded small unilamellar vesicles.

作者信息

Amselem S, Loyter A, Lichtenberg D, Barenholz Y

出版信息

Biochim Biophys Acta. 1985 Oct 24;820(1):1-10. doi: 10.1016/0005-2736(85)90208-1.

Abstract

The interaction of Sendai virus with small, unilamellar vesicles, lacking virus receptors and loaded with self-quenched 6-carboxyfluorescein, was studied. Sendai virions induced release of carboxyfluorescein from vesicles composed of negative charged phospholipids, despite the fact that they did not contain virus receptors. Preliminary experiments indicate that the carboxyfluorescein release is accompanied by mixing of the virus and liposome lipids and their entrapped contents, suggesting liposome-virus fusion. No release of carboxyfluorescein was observed with vesicles containing only phosphatidylcholine. The rate of virus-induced carboxyfluorescein release was temperature dependent; the lytic activity of the virus was greatly enhanced above 25 degrees C. This effect was not due to a thermal phase transition of the lipids in either the lipid vesicles or the virions. Virus-induced carboxyfluorescein release was inhibited by the presence of calcium ions in the medium and of cholesterol in the lipid vesicles. It increased with increasing concentrations of either the lipid vesicles or the virions. pretreatment of virions with increasing concentrations of three different proteolytic enzymes (trypsin, chymotrypsin and proteinase) inhibited the virus' ability to cause release of carboxyfluorescein from negatively charged liposomes. Inhibition of the viral lytic activity was also observed after virions were incubated above 56 degrees C.

摘要

研究了仙台病毒与缺乏病毒受体且装载有自猝灭6 - 羧基荧光素的小单层囊泡之间的相互作用。尽管不含有病毒受体,但仙台病毒粒子仍能诱导带负电荷磷脂组成的囊泡释放羧基荧光素。初步实验表明,羧基荧光素的释放伴随着病毒与脂质体脂质及其包裹内容物的混合,提示脂质体 - 病毒融合。仅含磷脂酰胆碱的囊泡未观察到羧基荧光素的释放。病毒诱导的羧基荧光素释放速率与温度有关;在25℃以上病毒的裂解活性大大增强。这种效应并非由于脂质体或病毒粒子中脂质的热相变所致。培养基中的钙离子和脂质体中的胆固醇会抑制病毒诱导的羧基荧光素释放。其释放随脂质体或病毒粒子浓度的增加而增加。用三种不同的蛋白水解酶(胰蛋白酶、胰凝乳蛋白酶和蛋白酶)以递增浓度预处理病毒粒子,会抑制病毒从带负电荷脂质体中引起羧基荧光素释放的能力。在病毒粒子于56℃以上孵育后,也观察到病毒裂解活性受到抑制。

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