Taguchi T, Kasai M
Biochim Biophys Acta. 1983 Apr 6;729(2):229-36. doi: 10.1016/0005-2736(83)90489-3.
This paper describes a new method of forming membrane vesicles. It was found that the addition of salt such as KCl into a solution containing lipid (asolectin) and a non-ionic surfactant, Triton X-114, led to the formation of closed membrane vesicles. The vesicles were separated from Triton X-114 by hydrophobic interaction chromatography. Electron microscopy revealed that the mean diameter of the vesicles was 110 nm +/- 69 nm (S.D.). Measurement of osmotic volume change showed that the permeability of the vesicle was very low to salts, sugar (glucose) and amphoteric ion (glycine), but very high to glycerol, ethylene glycol and water. Vesicle formation by this 'salting-out' method is very useful for reconstitution of transport systems in biomembranes because of its advantages: completion within a short time; high yield; and the possibility of utilizing samples in non-ionic surfactant solution. When we applied the method to the reconstitution of sarcoplasmic reticulum, Ca2+-ATPase was incorporated into the reconstituted vesicles and was enzymatically active in the membrane.
本文描述了一种形成膜泡的新方法。研究发现，向含有脂质（大豆卵磷脂）和非离子表面活性剂Triton X-114的溶液中添加诸如KCl的盐会导致形成封闭的膜泡。通过疏水相互作用色谱法将膜泡与Triton X-114分离。电子显微镜显示，膜泡的平均直径为110 nm±69 nm（标准差）。渗透体积变化的测量表明，膜泡对盐、糖（葡萄糖）和两性离子（甘氨酸）的通透性非常低，但对甘油、乙二醇和水的通透性非常高。由于这种“盐析”方法具有以下优点：在短时间内完成；产率高；以及能够利用非离子表面活性剂溶液中的样品，因此该方法对于生物膜中运输系统的重构非常有用。当我们将该方法应用于肌质网的重构时，Ca2+-ATP酶被整合到重构的膜泡中，并在膜中具有酶活性。
