Preparation of giant unilamellar vesicles from damp lipid film for better lipid compositional uniformity

被引:43
作者
Baykal-Caglar, Eda [1 ]
Hassan-Zadeh, Ebrahim [1 ]
Saremi, Bahar [1 ]
Huang, Juyang [1 ]
机构
[1] Texas Tech Univ, Dept Phys, Lubbock, TX 79409 USA
来源
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES | 2012年 / 1818卷 / 11期
关键词
Miscibility transition temperature; Electroformation; Cholesterol; Umbrella Model; Lipid demixing; LATERAL DISTRIBUTION; MAXIMUM SOLUBILITY; TERNARY MIXTURES; PHASE-BEHAVIOR; CHOLESTEROL; MEMBRANES; MODEL; LIPOSOMES;
D O I
10.1016/j.bbamem.2012.05.023
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
Giant unilamellar vesicles (GUVs) containing cholesterol often have a wide distribution in lipid composition. In this study, GUVs of 1,2-dioleoyl-sn-glycero-3-phosphocholine(DOPC)/1,2-distearoyl-sn-glycero-3-phosphocholine(DSPC)/cholesterol and 1,2-diphytanoyl-sn-glycero-3-phosphocholine(diPhyPC)/1,2-dipalmitoyl-sn-glycero-3-phosphocholine(DPPC)/cholesterol were prepared from dry lipid films using the standard electroformation method as well as a modified method from damp lipid films, which are made from compositional uniform liposomes prepared using the Rapid Solvent Exchange (RSE) method. We quantified the lipid compositional distributions of GUV by measuring the miscibility transition temperature of GUVs using fluorescence microscopy, since a narrower distribution in the transition temperature should correspond to a more uniform distribution in GIN lipid composition. Cholesterol molecules can demix from other lipids in dry state and form cholesterol crystals. Using optical microscopy, micron-sized crystals were observed in some dry lipid films. Thus, a major cause of GUV lipid compositional heterogeneity is the demixing of lipids in the dry film state. By avoiding the dry film state, GUVs prepared from damp lipid films have a better uniformity in lipid composition, and the standard deviations of miscibility transition temperature are about 2.5 times smaller than that of GUVs prepared from dry lipid films. Comparing the two ternary systems, diPhyPC/DPPC/cholesterol GUVs has a larger cholesterol compositional heterogeneity, which directly correlates with the low maximum solubility of cholesterol in diPhyPC lipid bilayers (40.2 +/- 0.5 mol%) measured by light scattering. Our data indicate that cholesterol interacts far less favorably with diPhyPC than it does with other PCs. The damp lipid film method also has a potential of preparing GUVs from cell membranes containing native proteins without going through a dry state. (C) 2012 Elsevier B.V. All rights reserved.
引用
收藏
页码:2598 / 2604
页数:7
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