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聚天冬氨酸依赖的携带季铵洗涤剂[[[(1,1,3,3 - 四甲基丁基)甲酚氧基]乙氧基]乙基]二甲基苄基氢氧化铵的脂质体融合。

Poly(aspartic acid)-dependent fusion of liposomes bearing the quaternary ammonium detergent [[[(1,1,3,3-tetramethylbutyl)cresoxy]ethoxy]ethyl] dimethylbenzylammonium hydroxide.

作者信息

Beigel M, Keren-Zur M, Laster Y, Loyter A

机构信息

International Genetic Sciences Partnership, Jerusalem, Israel.

出版信息

Biochemistry. 1988 Jan 26;27(2):660-6. doi: 10.1021/bi00402a025.

Abstract

Addition of the quaternary ammonium detergent [[[(1,1,3,3-tetramethylbutyl)cresoxy]ethoxy]ethyl] dimethylbenzylammonium hydroxide (DEBDA[OH]) and the fluorescent probes N-(7-nitro-2-1,3-benzoxadiazol-4-yl)phosphatidylethanolamine and N-(lissamine rhodamine B sulfonyl)phosphatidylethanolamine (N-NBD-PE and N-Rh-PE, respectively) to liposomes composed of phosphatidylcholine (PC) and cholesterol (chol) resulted in the formation of fluorescently labeled liposomes bearing DEBDA[OH]. Incubation of the anionic polymer poly(aspartic acid) (PASP) with such liposomes resulted in strong agglutination, indicating an association between the negatively charged PASP and the positively charged liposome-associated DEBDA[OH]. Addition of PASP to a mixture of fluorescently labeled and nonlabeled liposomes, both carrying DEBDA[OH], resulted in a significant increase in the extent of fluorescence, namely, fluorescence dequenching. The degree of the fluorescence dequenching was dependent upon the ratio between the nonfluorescent and the fluorescent liposomes, upon the temperature of incubation, and upon the amount of DEBDA[OH] which was associated with the liposomes. Electron microscopic observations revealed that large liposomes were formed upon incubation of liposomes bearing DEBDA[OH] with PASP. The results of the present work strongly indicate that the fluorescence dequenching observed is due to a process of PASP-induced liposome-liposome fusion.

摘要

向由磷脂酰胆碱(PC)和胆固醇(chol)组成的脂质体中添加季铵洗涤剂[[[(1,1,3,3 - 四甲基丁基)甲酚氧基]乙氧基]乙基]二甲基苄基氢氧化铵(DEBDA[OH])以及荧光探针N - (7 - 硝基 - 2 - 1,3 - 苯并恶二唑 - 4 - 基)磷脂酰乙醇胺和N - (丽丝胺罗丹明B磺酰基)磷脂酰乙醇胺(分别为N - NBD - PE和N - Rh - PE),导致形成带有DEBDA[OH]的荧光标记脂质体。将阴离子聚合物聚天冬氨酸(PASP)与这种脂质体一起孵育会导致强烈凝集,表明带负电荷的PASP与带正电荷的脂质体相关的DEBDA[OH]之间存在缔合。向均带有DEBDA[OH]的荧光标记和未标记脂质体的混合物中添加PASP,导致荧光程度显著增加,即荧光猝灭解除。荧光猝灭解除的程度取决于非荧光脂质体与荧光脂质体的比例、孵育温度以及与脂质体相关的DEBDA[OH]的量。电子显微镜观察表明,将带有DEBDA[OH]的脂质体与PASP一起孵育时会形成大的脂质体。本研究结果有力地表明,观察到的荧光猝灭解除是由于PASP诱导的脂质体 - 脂质体融合过程。

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