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    题名: 麥角固醇對含膽固醇的脂雙層膜的影響
    作者: 許家碩;Hsu,Chia Shuo
    贡献者: 生物物理研究所
    关键词: 麥角固醇;膽固醇;核磁共振儀;人造細胞膜;NMR;POPC;DPPC;ergosterol;cholesterol
    日期: 2015-07-28
    上传时间: 2015-09-23 10:53:24 (UTC+8)
    出版者: 國立中央大學
    摘要: 膽固醇(cholesterol)和麥角固醇(ergosterol)的結構相似,然而有些研究指出膽固醇和麥角固醇對脂雙層膜的影響不同。在有些細胞膜中發現同時含有膽固醇及麥角固醇,如釀酒酵母菌的粒腺體。因此我們想了解含有膽固醇與麥角固醇的脂雙層膜的特性為何。我們利用核磁共振(2H NMR)以及螢光顯微術來研究不飽和脂質、飽和脂質、膽固醇以及麥角固醇所組成人造膜的物理性質。核磁共振則藉由氘化不飽和脂質分子的sn-1碳氫鏈,得以測量脂質膜的相行為和脂質膜的有序程度。螢光顯微術藉由螢光分子(NBD-DOPE)標示液態無序相(liquid disorder, ld),以探討人造膜的表面形態。我們固定不飽和/飽和脂質的莫耳數比改變膽固醇/麥角固醇的莫耳數比作一系列的量測。我們也改變溫度進行一系列的量測。觀察到在相同溫度下,含膽固醇的脂雙層膜處於 lo 態,加入ergosterol會促進ld態的產生,增加ergosterol的含量,ld態含量增加,使得脂雙層膜有序程度減少。對於每種脂質膜,溫度升高,也會使脂雙層膜有序程度減少。;The structure of cholesterol and ergosterol are similar, whereas the effect of cholesterol and ergosterol on lipid bilayer is different. Some cell membranes contain both cholesterol and ergosterol, such as the mitochondria of Saccharomyces cerevisiae. How ergosterol affects the properties cholesterol-containing lipid bilayers is of interest. In this work, we have studied the physical properties of model membranes containing unsaturated lipid, saturated lipid, cholesterol, and ergosterol using deuterium nuclear magnetic resonance (2H-NMR) and fluorescence microscopy.In the NMR experiment, unsaturated lipids were deuterium labeled. The phase behavior and membrane order then can be observed.The morphology of model membranes was observed by fluorescence microscopy. The fluorescent probe NBD-DOPE, presumably participate into the disordered phases, is used.The measurements were performed as a function of cholesterol/ergosterol molar ratio by fixing the molar ratio of unsaturated to saturated lipids.The measurements were also performed as a function of temperature. Addition of ergosterol to the cholesterol-containing lipid bilayers, which is in the lo phase for a given temperature,produces the ld phase. The fraction of the ld phase increases as the ergosterol increases, resulting in the decrease of membrane order. Furthermore, it is observed that for each membrane composition, the membrane order decreases with increasing temperature.
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