Hernandez Victor Agmo, Scholz Fritz
Institut für Biochemie, Universität Greifswald, 17489 Greifswald, Soldmannstr. 23, Germany.
Langmuir. 2006 Dec 5;22(25):10723-31. doi: 10.1021/la060908o.
Liposomes suspended in aqueous electrolyte solutions can adhere at mercury electrodes. The adhesion is a complex process that starts with the docking and opening and leads to a spreading, finally resulting in the formation of islands of adsorbed lecithin molecules. The adhesion process can be followed by chronoamperometry, and a detailed analysis of the macroscopic and microscopic kinetics can be performed yielding rate constants and activation parameters. By using giant unilamellar liposomes and multilamellar liposomes, the effect of lamellarity and liposome size could be elucidated for liposomes in the liquid crystalline, gel, and superlattice phase states. Below the phase transition temperature, the time constant of opening of the liposomes (i.e., the irreversible binding of the lecithin molecules on the preliminary contact interface liposome|mercury and the therewith associated disintegration of the liposome membrane on that spot) is shown to be strongly size dependent. The activation energy, however, of that process is size independent with the exception of very small liposomes. That size dependence of time constants is a result of the size dependence of the initial contact area. The time constant and the activation energies of the spreading step exhibit a strong size dependence, which could be shown to be due to the size dependence of rate and activation energy of pore formation. Pore formation is necessary to release the solution included in the liposomes. This understanding was corroborated by addition of the pore inducing peptide Mastoparan X to the liposome suspension. The obtained results show that electrochemical studies of liposome adhesion on mercury electrodes can be used as a biomimetic tool to understand the effect of membrane properties on vesicle fusion.
悬浮于电解质水溶液中的脂质体可粘附在汞电极上。这种粘附是一个复杂的过程,始于对接与开口,进而发展为铺展,最终导致吸附的卵磷脂分子形成岛状结构。粘附过程可通过计时电流法进行跟踪,并且可以对宏观和微观动力学进行详细分析,从而得出速率常数和活化参数。通过使用巨型单层脂质体和多层脂质体,可以阐明层状结构和脂质体大小对处于液晶、凝胶和超晶格相态的脂质体的影响。在相变温度以下,脂质体开口的时间常数(即卵磷脂分子在脂质体|汞的初步接触界面上的不可逆结合以及脂质体膜在该位置的相关崩解)显示出强烈的尺寸依赖性。然而,除了非常小的脂质体之外,该过程的活化能与尺寸无关。时间常数的这种尺寸依赖性是初始接触面积尺寸依赖性的结果。铺展步骤的时间常数和活化能表现出强烈的尺寸依赖性,这可以证明是由于孔形成速率和活化能的尺寸依赖性所致。孔形成对于释放脂质体中包含的溶液是必要的。通过向脂质体悬浮液中添加诱导孔形成的肽马蜂毒素X,证实了这一认识。所得结果表明,脂质体在汞电极上粘附的电化学研究可作为一种仿生工具,用于理解膜性质对囊泡融合的影响。