Stein Hannah, Spindler Susann, Bonakdar Navid, Wang Chun, Sandoghdar Vahid
Friedrich-Alexander University Erlangen-NurembergErlangen, Germany; Max Planck Institute for the Science of LightErlangen, Germany.
Max Planck Institute for the Science of Light Erlangen, Germany.
Front Physiol. 2017 Feb 13;8:63. doi: 10.3389/fphys.2017.00063. eCollection 2017.
The cell membrane forms a dynamic and complex barrier between the living cell and its environment. However, its studies are difficult because it consists of a high variety of lipids and proteins and is continuously reorganized by the cell. Therefore, membrane model systems with precisely controlled composition are used to investigate fundamental interactions of membrane components under well-defined conditions. Giant unilamellar vesicles (GUVs) offer a powerful model system for the cell membrane, but many previous studies have been performed in unphysiologically low ionic strength solutions which might lead to altered membrane properties, protein stability and lipid-protein interaction. In the present work, we give an overview of the existing methods for GUV production and present our efforts on forming single, free floating vesicles up to several tens of μm in diameter and at high yield in various buffer solutions with physiological ionic strength and pH.
细胞膜在活细胞与其环境之间形成了一个动态且复杂的屏障。然而,对其进行研究颇具难度,因为它由种类繁多的脂质和蛋白质组成,且细胞会不断对其进行重组。因此,具有精确可控组成的膜模型系统被用于研究在明确条件下膜成分的基本相互作用。巨型单层囊泡(GUVs)为细胞膜提供了一个强大的模型系统,但此前许多研究是在非生理条件下的低离子强度溶液中进行的,这可能会导致膜性质、蛋白质稳定性以及脂质 - 蛋白质相互作用发生改变。在本研究中,我们概述了现有的GUV制备方法,并展示了我们在各种具有生理离子强度和pH值的缓冲溶液中,以高产量形成直径达数十微米的单个、自由漂浮囊泡的成果。