Department of Chemistry, The Scripps Research Institute, 130 Scripps Way, Jupiter, Florida 33458, USA.
Nat Chem. 2013 Nov;5(11):958-63. doi: 10.1038/nchem.1765. Epub 2013 Sep 29.
Eukaryotic subcellular membrane systems, such as the nuclear envelope or endoplasmic reticulum, present a rich array of architecturally and compositionally complex supramolecular targets that are as yet inaccessible. Here we describe layer-by-layer phospholipid membrane assembly on microfluidic droplets, a route to structures with defined compositional asymmetry and lamellarity. Starting with phospholipid-stabilized water-in-oil droplets trapped in a static droplet array, lipid monolayer deposition proceeds as oil/water-phase boundaries pass over the droplets. Unilamellar vesicles assembled layer-by-layer support functional insertion both of purified and of in situ expressed membrane proteins. Synthesis and chemical probing of asymmetric unilamellar and double-bilayer vesicles demonstrate the programmability of both membrane lamellarity and lipid-leaflet composition during assembly. The immobilized vesicle arrays are a pragmatic experimental platform for biophysical studies of membranes and their associated proteins, particularly complexes that assemble and function in multilamellar contexts in vivo.
真核亚细胞膜系统，如核膜或内质网，呈现出丰富多样的结构和组成上复杂的超分子靶标，这些靶标目前还无法接近。在这里，我们描述了在微流控液滴上的层层磷脂膜组装，这是一种具有明确组成不对称性和层状结构的途径。从在静态液滴阵列中捕获的磷脂稳定的油包水液滴开始，当油/水相边界越过液滴时，脂质单层沉积就会进行。通过层层组装形成的单层囊泡支持纯化的和原位表达的膜蛋白的功能插入。不对称的单层和双层囊泡的合成和化学探测证明了在组装过程中膜的层状结构和脂质叶层组成的可编程性。固定化囊泡阵列是用于膜及其相关蛋白的生物物理研究的实用实验平台，特别是在体内以多层层状结构组装和发挥功能的复合物。
