Cholesterol-dependent phase separation in cell-derived giant plasma-membrane vesicles

被引：127

作者：

Levental, Ilya ^{[2
]}

Byfield, Fitzroy J. ^{[3
]}

Chowdhury, Pramit ^{[4
]}

Gai, Feng ^{[4
]}

Baumgart, Tobias ^{[4
]}

Janmey, Paul A. ^{[1
,5
]}

机构：

[1] Univ Penn, Dept Phys, Vagelos Labs 1010, Philadelphia, PA 19104 USA

[2] Univ Penn, Dept Bioengn, Philadelphia, PA 19104 USA

[3] Univ Penn, Inst Med & Engn, Vagelos Labs 1010, Philadelphia, PA 19104 USA

[4] Univ Penn, Dept Chem, Vagelos Labs 1010, Philadelphia, PA 19104 USA

[5] Univ Penn, Dept Physiol, Vagelos Labs 1010, Philadelphia, PA 19104 USA

来源：

BIOCHEMICAL JOURNAL | 2009年 / 424卷

基金：

美国国家卫生研究院; 美国国家科学基金会;

关键词：

giant plasma-membrane vesicle (GPMV); lipid raft; liquid ordered; FLUORESCENCE CORRELATION SPECTROSCOPY; GPI-ANCHORED PROTEINS; LIPID RAFTS; MODEL MEMBRANES; PHOSPHOLIPIDS; DOMAINS; TEMPERATURE; MICROSCOPY; MONOLAYERS; MIXTURES;

D O I：

10.1042/BJ20091283

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Cell-derived GPMVs (giant plasma-membrane vesicles) enable investigation of lipid phase separation in a system with appropriate biological complexity under physiological conditions, and in the present study were used to investigate the cholesterol-dependence of domain formation and stability. The cholesterol level is directly related to the abundance of the liquid-ordered phase fraction, which is the majority phase in vesicles from untreated cells. Miscibility transition temperature depends on cholesterol and correlates strongly with the presence of detergent-insoluble membrane in cell lysates. Fluorescence correlation spectroscopy reveals two distinct diffusing populations in phase-separated cell membrane-derived vesicles whose diffusivities correspond well to diffusivities in both model systems and live cells. The results of the present study extend previous observations in purified lipid systems to the complex environment of the plasma membrane and provide insight into the effect of cholesterol on lipid phase separation and abundance.

引用

页码：163 / 167

页数：5

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