Biofisika Institute (UPV/EHU, CSIC) and Department of Biochemistry and Molecular Biology, University of the Basque Country, Bilbao, Spain.
Sci Rep. 2019 May 10;9(1):7255. doi: 10.1038/s41598-019-43561-4.
In vitro reconstitution and microscopic visualization of membrane processes is an indispensable source of information about a cellular function. Here we describe a conceptionally novel free-standing membrane template that facilitates such quantitative reconstitution of membrane remodelling at different scales. The Giant Suspended Bilayers (GSBs) spontaneously swell from lipid lamella reservoir deposited on microspheres. GSBs attached to the reservoir can be prepared from virtually any lipid composition following a fast procedure. Giant unilamellar vesicles can be further obtained by GSB detachment from the microspheres. The reservoir stabilizes GSB during deformations, mechanical micromanipulations, and fluorescence microscopy observations, while GSB-reservoir boundary enables the exchange of small solutes with GSB interior. These unique properties allow studying macro- and nano-scale membrane deformations, adding membrane-active compounds to both sides of GSB membrane and applying patch-clamp based approaches, thus making GSB a versatile tool for reconstitution and quantification of cellular membrane trafficking events.
体外重建和微观可视化膜过程是了解细胞功能的不可或缺的信息来源。在这里，我们描述了一种新颖的独立式膜模板，可促进不同尺度的膜重塑的定量重建。巨悬浮双层膜（GSB）自发地从沉积在微球上的脂质双层储库中膨胀。GSB 可以从几乎任何脂质组成中通过快速程序附着在储库上制备。通过从微球上分离 GSB 可以进一步获得巨大的单层囊泡。储库在变形、机械微操作和荧光显微镜观察过程中稳定 GSB，而 GSB-储库边界允许与 GSB 内部交换小分子溶质。这些独特的特性允许研究宏观和纳米尺度的膜变形，在 GSB 膜的两侧添加膜活性化合物，并应用基于膜片钳的方法，从而使 GSB 成为重建和量化细胞膜运输事件的多功能工具。
