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关于两种平面脂质双层——折叠型和涂抹型——与囊泡融合的定量比较。

Quantitative comparison of two types of planar lipid bilayers--folded and painted--with respect to fusion with vesicles.

作者信息

Anzai K, Ogawa K, Ozawa T, Yamamoto H

机构信息

Bioregulation Research Group, National Institute of Radiological Sciences, 4-9-1 Anagawa, Chiba 263-8555, Inage, Japan.

出版信息

J Biochem Biophys Methods. 2001 May 28;48(3):283-91. doi: 10.1016/s0165-022x(01)00160-9.

Abstract

Two major types of planar lipid bilayers, painted and folded, were compared with respect to vesicle fusion using one chamber for the preparation of both bilayers. Liposomes containing ion channels composed of nystatin and ergosterol were used as the vesicle sample. Fusion of the liposome to either bilayer elicited a spike-like current change, which corresponds to a fusion event. The lag time between the first fusion event and the addition of the vesicles is an index of the ease with which the vesicles fuse with the bilayers. The lag time in the painted bilayer at a KCl concentration (cis) of 450 mM was 1.58+/-1.18 min, similar to that in the folded bilayer (1.65+/-0.64 min). The lag time decreased with increase of the osmotic difference across the painted bilayer, whereas this change was small in the folded bilayer. The fusion of the liposomes to the painted bilayer was markedly reduced by stopping the stirring in the cis compartment, whereas the fusion to the folded bilayer was not affected significantly. These results imply that no practical difference exists in the ability of vesicles to fuse with the painted and folded bilayers. For the study of single channel behavior, the painted bilayer could have an advantage because simply stopping the stirring prevents excess fusion.

摘要

使用一个腔室来制备两种平面脂质双层(涂抹型和折叠型),比较了这两种主要类型的平面脂质双层在囊泡融合方面的情况。含有由制霉菌素和麦角固醇组成的离子通道的脂质体用作囊泡样本。脂质体与任一脂质双层的融合引发了尖峰状电流变化,这对应于一个融合事件。第一次融合事件与添加囊泡之间的滞后时间是囊泡与脂质双层融合难易程度的一个指标。在450 mM的KCl浓度(顺式)下,涂抹型脂质双层中的滞后时间为1.58±1.18分钟,与折叠型脂质双层中的滞后时间(1.65±0.64分钟)相似。滞后时间随着涂抹型脂质双层两侧渗透压差的增加而减少,而在折叠型脂质双层中这种变化很小。通过停止顺式隔室中的搅拌,脂质体与涂抹型脂质双层的融合明显减少,而与折叠型脂质双层的融合没有受到显著影响。这些结果表明,囊泡与涂抹型和折叠型脂质双层融合的能力没有实际差异。对于单通道行为的研究,涂抹型脂质双层可能具有优势,因为简单地停止搅拌可以防止过度融合。

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