Veatch Sarah L, Keller Sarah L
Departments of Chemistry and Physics, University of Washington, Seattle, Washington 98195-1700, USA.
Biophys J. 2003 Nov;85(5):3074-83. doi: 10.1016/S0006-3495(03)74726-2.
We use fluorescence microscopy to directly observe liquid phases in giant unilamellar vesicles. We find that a long list of ternary mixtures of high melting temperature (saturated) lipids, low melting temperature (usually unsaturated) lipids, and cholesterol produce liquid domains. For one model mixture in particular, DPPC/DOPC/Chol, we have mapped phase boundaries for the full ternary system. For this mixture we observe two coexisting liquid phases over a wide range of lipid composition and temperature, with one phase rich in the unsaturated lipid and the other rich in the saturated lipid and cholesterol. We find a simple relationship between chain melting temperature and miscibility transition temperature that holds for both phosphatidylcholine and sphingomyelin lipids. We experimentally cross miscibility boundaries both by changing temperature and by the depletion of cholesterol with beta-cyclodextrin. Liquid domains in vesicles exhibit interesting behavior: they collide and coalesce, can finger into stripes, and can bulge out of the vesicle. To date, we have not observed macroscopic separation of liquid phases in only binary lipid mixtures.
我们使用荧光显微镜直接观察巨型单层囊泡中的液相。我们发现,一长串由高熔点(饱和)脂质、低熔点(通常为不饱和)脂质和胆固醇组成的三元混合物会产生液相区域。特别是对于一种模型混合物DPPC/DOPC/Chol,我们绘制了整个三元体系的相界。对于这种混合物,我们在很宽的脂质组成和温度范围内观察到两个共存的液相,一个相富含不饱和脂质,另一个相富含饱和脂质和胆固醇。我们发现链熔化温度和混溶转变温度之间存在一种简单的关系,这种关系适用于磷脂酰胆碱和鞘磷脂。我们通过改变温度以及用β-环糊精消耗胆固醇来实验性地跨越混溶边界。囊泡中的液相区域表现出有趣的行为:它们会碰撞并合并,能形成指状条纹,还能从囊泡中鼓出。迄今为止,我们尚未在仅由二元脂质混合物中观察到液相的宏观分离。