Siegel D P, Tenchov B G
Givaudan Inc., Cincinnati, Ohio 45216, USA.
Biophys J. 2008 May 15;94(10):3987-95. doi: 10.1529/biophysj.107.118034. Epub 2008 Jan 30.
Based on curvature energy considerations, nonbilayer phase-forming phospholipids in excess water should form stable bicontinuous inverted cubic (Q(II)) phases at temperatures between the lamellar (L(alpha)) and inverted hexagonal (H(II)) phase regions. However, the phosphatidylethanolamines (PEs), which are a common class of biomembrane phospholipids, typically display direct L(alpha)/H(II) phase transitions and may form intermediate Q(II) phases only after the temperature is cycled repeatedly across the L(alpha)/H(II) phase transition temperature, T(H), or when the H(II) phases are cooled from T > T(H). This raises the question of whether models of inverted phase stability, which are based on curvature energy alone, accurately predict the relative free energy of these phases. Here we demonstrate the important role of a noncurvature energy contribution, the unbinding energy of the L(alpha) phase bilayers, g(u), that serves to stabilize the L(alpha) phase relative to the nonlamellar phases. The planar L(alpha) phase bilayers must separate for a Q(II) phase to form and it turns out that the work of their unbinding can be larger than the curvature energy reduction on formation of Q(II) phase from L(alpha) at temperatures near the L(alpha)/Q(II) transition temperature (T(Q)). Using g(u) and elastic constant values typical of unsaturated PEs, we show that g(u) is sufficient to make T(Q) > T(H) for the latter lipids. Such systems would display direct L(alpha) --> H(II) transitions, and a Q(II) phase might only form as a metastable phase upon cooling of the H(II) phase. The g(u) values for methylated PEs and PE/phosphatidylcholine mixtures are significantly smaller than those for PEs and increase T(Q) by only a few degrees, consistent with observations of these systems. This influence of g(u) also rationalizes the effect of some aqueous solutes to increase the rate of Q(II) formation during temperature cycling of lipid dispersions. Finally, the results are relevant to protocols for determining the Gaussian curvature modulus, which substantially affects the energy of intermediates in membrane fusion and fission. Recently, two such methods were proposed based on measuring T(Q) and on measuring Q(II) phase unit cell dimensions, respectively. In view of the effect of g(u) on T(Q) that we describe here, the latter method, which does not depend on the value of g(u), is preferable.
基于曲率能量的考虑,在过量水中,形成非双层相的磷脂在层状(L(α))相和反六角(H(II))相区域之间的温度下应形成稳定的双连续反立方(Q(II))相。然而,磷脂酰乙醇胺(PEs)作为生物膜磷脂的常见类别,通常呈现直接的L(α)/H(II)相变,并且可能仅在温度反复循环通过L(α)/H(II)相变温度T(H)之后,或者当H(II)相从T > T(H)冷却时,才形成中间的Q(II)相。这就引发了一个问题,即仅基于曲率能量的反相稳定性模型是否能准确预测这些相的相对自由能。在此,我们证明了一种非曲率能量贡献——L(α)相双层的解束缚能g(u)的重要作用,它相对于非层状相起到稳定L(α)相的作用。平面L(α)相双层必须分离才能形成Q(II)相,结果表明,在接近L(α)/Q(II)转变温度(T(Q))时,它们解束缚的功可能大于从L(α)形成Q(II)相时曲率能量的降低。使用不饱和PEs典型的g(u)和弹性常数数值,我们表明g(u)足以使后者脂质的T(Q) > T(H)。这样的系统将呈现直接的L(α)→H(II)转变,并且Q(II)相可能仅在H(II)相冷却时作为亚稳相形成。甲基化PEs和PE/磷脂酰胆碱混合物的g(u)值明显小于PEs的g(u)值,并且仅使T(Q)升高几度,这与这些系统的观察结果一致。g(u)的这种影响也解释了一些水性溶质在脂质分散体温度循环过程中增加Q(II)形成速率的作用。最后,这些结果与确定高斯曲率模量的方案相关,高斯曲率模量会显著影响膜融合和裂变中间体的能量。最近,分别基于测量T(Q)和测量Q(II)相晶胞尺寸提出了两种这样的方法。鉴于我们在此描述的g(u)对T(Q)的影响,后一种不依赖于g(u)值的方法更可取。