Haines T H, Li W, Green M, Cummins H Z
Department of Chemistry, City College of The City University of New York, New York 10031.
Biochemistry. 1987 Aug 25;26(17):5439-47. doi: 10.1021/bi00391a034.
Uniform, unilamellar vesicles have been prepared by the pH-modification technique. The initial sizes of the vesicles were from 200 to 700 nm and were measured to within 1-3% by photo correlation spectroscopy. Vesicles were made of the dioleoyl esters of phosphatidic acid, phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, the diphytanyl ethers of phosphatidylglycerol, Escherichia coli lipids, and lac permease reconstituted into E. coli lipids. The vesicle suspensions were prepared and then diluted with electrolyte (KCl) and/or nonelectrolyte (sucrose, trehalose, pentaerythritol) impermeants. The amplitude of the swelling is linearly proportional to the osmotic pressure difference across the bilayer. We have determined the elastic modulus, the elastic limit (percent surface expansion at bursting), and the transbilayer pressure difference at bursting for each of these vesicles at constant osmolarity but at different ionic strengths. We find that the elastic properties of the bilayer vary by a factor of 10 in electrolyte media as compared to isosmolal nonelectrolyte media and that this variation appears to be related to both the charge density at the surface and the ionic strength of the media. Anionic lipid vesicles in 150 mM KCl have a significantly higher modulus (50 X 10(7) dyn/cm2) and transbilayer pressure difference (40 mosM) at bursting with a small capacity to stretch (3-4% surface expansion) compared to the same vesicles suspended in nonelectrolyte impermeants. The latter vesicles undergo a significant surface expansion (8-10%), display a low modulus (3 X 10(7) dyn/cm2), and burst at 3-4 mosM bilayer pressure difference. Vesicles suspended in media of constant osmolarity at various ionic strengths display properties with proportional values.(ABSTRACT TRUNCATED AT 250 WORDS)
通过pH调节技术制备了均匀的单层囊泡。囊泡的初始大小为200至700纳米,通过光散射光谱法测量其误差在1%-3%以内。囊泡由磷脂酸、磷脂酰胆碱、磷脂酰甘油、磷脂酰乙醇胺的二油酰酯、磷脂酰甘油的二植烷基醚、大肠杆菌脂质以及重组到大肠杆菌脂质中的乳糖通透酶制成。制备囊泡悬浮液,然后用电解质(氯化钾)和/或非电解质(蔗糖、海藻糖、季戊四醇)等非渗透性物质进行稀释。肿胀幅度与跨双层的渗透压差值呈线性比例关系。我们在恒定渗透压但不同离子强度下,测定了每种囊泡的弹性模量、弹性极限(破裂时的表面膨胀百分比)以及破裂时的跨双层压差。我们发现,与等渗非电解质介质相比,双层膜的弹性特性在电解质介质中变化了10倍,这种变化似乎与表面电荷密度和介质的离子强度都有关。与悬浮在非电解质非渗透性物质中的相同囊泡相比,在150 mM氯化钾中的阴离子脂质囊泡在破裂时具有显著更高的模量(50×10⁷ 达因/平方厘米)和跨双层压差(40 mosM),且拉伸能力较小(表面膨胀3%-4%)。后者的囊泡会发生显著的表面膨胀(8%-10%),显示出低模量(3×10⁷ 达因/平方厘米),并在3-4 mosM的双层压差下破裂。悬浮在不同离子强度的恒定渗透压介质中的囊泡显示出具有比例值的特性。(摘要截断于250字)