Biner Olivier, Schick Thomas, Müller Yannic, von Ballmoos Christoph
Department of Chemistry and Biochemistry, University of Bern, Switzerland.
Graduate School for Cellular and Biomedical Sciences, University of Bern, Switzerland.
FEBS Lett. 2016 Jul;590(14):2051-62. doi: 10.1002/1873-3468.12233. Epub 2016 Jun 21.
One of the current challenges in synthetic biology is the production of stable membrane mimetic systems and the insertion of components in these systems. Here, we employ fusion of oppositely charged liposomes to deliver separately reconstituted membrane proteins into a common lipid bilayer. After a systematic evaluation of different lipid compositions by lipid mixing and size distribution analysis, suitable conditions were further investigated for proteoliposome fusion. With this technique, we functionally coreconstituted bo3 oxidase and ATP synthase from Escherichia coli into unilamellar liposomes ranging from 100 nm to 50 μm in size. The presented method is a simple and versatile tool for oriented membrane protein reconstitution to produce biomimetic systems with increased complexity.
合成生物学当前面临的挑战之一是构建稳定的膜模拟系统并将组件插入这些系统。在此，我们利用带相反电荷的脂质体融合，将分别重构的膜蛋白递送至共同的脂质双层中。通过脂质混合和尺寸分布分析对不同脂质组成进行系统评估后，进一步研究了适合蛋白脂质体融合的条件。利用该技术，我们将来自大肠杆菌的bo3氧化酶和ATP合酶功能共重构到尺寸范围为100纳米至50微米的单层脂质体中。所提出的方法是一种简单且通用的工具，用于定向膜蛋白重构以产生具有更高复杂性的仿生系统。
