Needham D, McIntosh T J, Evans E
Department of Pathology, University of British Columbia, Vancouver, Canada.
Biochemistry. 1988 Jun 28;27(13):4668-73. doi: 10.1021/bi00413a013.
Mixtures of dimyristoylphosphatidylcholine (DMPC) and cholesterol (Chol) have been used to examine the effects of cholesterol on the chain crystallization transitions and thermomechanical properties in phospholipid bilayer membranes. The mechanical properties--elastic moduli and level of tension at membrane rupture--were derived from micropipet pressurization of giant single-walled vesicles. Also, the micropipet method allowed temperature-dependent area transitions to be measured at constant membrane tension. X-ray diffraction measurements were made on selected lipid/cholesterol mixtures. Wide-angle patterns and electron density profiles were used to measure bilayer thickness as an indication of chain tilt and fluidity. Vesicle area versus temperature plots showed that the main acyl chain crystallization transition of DMPC broadened and shifted to higher temperatures. Both above and below the broad transition, the elastic area compressibility modulus, K, was greatly increased with cholesterol addition. The value for the 1:1 DMPC/Chol complex was found to be approximately 700 dyn/cm, comparable to that for DMPC in the L beta' phase. However, for all concentrations above 12.5 mol % (which was weakly solid), vesicle bilayers behaved as surface liquids with no surface shear rigidity even at temperatures well below the DMPC phase transition. Area changes over the broadened transitions were reduced by cholesterol and disappeared with the addition of 50 mol % to leave the thermal area expansivity at 1.3 X 10(-3)/degrees C. These area changes are consistent with separate formation of a 1:1 DMPC/Chol complex that does not condense plus residual free lipid and lipid loosely associated with the 1:1 complex that freezes normally.(ABSTRACT TRUNCATED AT 250 WORDS)
二肉豆蔻酰磷脂酰胆碱(DMPC)与胆固醇(Chol)的混合物已被用于研究胆固醇对磷脂双层膜中链结晶转变和热机械性能的影响。通过对巨型单壁囊泡进行微量移液管加压得出其力学性能——弹性模量和膜破裂时的张力水平。此外,微量移液管方法还能在恒定膜张力下测量与温度相关的面积转变。对选定的脂质/胆固醇混合物进行了X射线衍射测量。利用广角图谱和电子密度分布图来测量双层膜厚度,以此作为链倾斜和流动性的指标。囊泡面积与温度的关系图表明,DMPC的主要酰基链结晶转变变宽并向更高温度移动。在宽转变温度之上和之下,添加胆固醇后弹性面积压缩模量K均大幅增加。发现1:1 DMPC/Chol复合物的值约为700达因/厘米,与Lβ'相中DMPC的值相当。然而,对于所有高于12.5摩尔%(呈弱固态)的浓度,即使在远低于DMPC相变温度的情况下,囊泡双层膜也表现为表面液体,没有表面剪切刚性。胆固醇减少了宽转变过程中的面积变化,添加50摩尔%胆固醇后面积变化消失,热面积膨胀系数为1.3×10⁻³/℃。这些面积变化与1:1 DMPC/Chol复合物的单独形成一致,该复合物不会凝聚,再加上残留的游离脂质以及与正常冻结的1:1复合物松散结合的脂质。(摘要截短于250字)