Univ. Bordeaux, CNRS, Bordeaux INP, LCPO, UMR 5629, 33600, Pessac, France.
Angew Chem Int Ed Engl. 2020 Jun 26;59(27):11028-11036. doi: 10.1002/anie.202001868. Epub 2020 Apr 28.
Elastin-like polypeptides (ELPs) have been proposed as a simple model of intrinsically disordered proteins (IDPs) which can form membraneless organelles by liquid-liquid phase separation (LLPS) in cells. Herein, the behavior of fluorescently labeled ELP is studied in cytomimetic aqueous two-phase system (ATPS) encapsulated protocells that are formed using microfluidics, which enabled confinement, changes in temperature, and statistical analysis. The spatial organization of ELP could be observed in the ATPS. Furthermore, changes in temperature triggered the dynamic formation and distribution of ELP-rich droplets within the ATPS, resulting from changes in conformation. Proteins were encapsulated along with ELP in the synthetic protocells and distinct partitioning properties of these proteins and ELP in the ATPS were observed. Therefore, the ability of ELP to coacervate with temperature can be maintained inside a cell-mimicking system.
弹性蛋白样多肽 (ELP) 被提出作为一种简单的无规卷曲蛋白质 (IDP) 模型,它可以通过细胞内的液-液相分离 (LLPS) 形成无膜细胞器。在此,通过使用微流控技术形成的细胞模拟水相双相体系 (ATPS) 中荧光标记的 ELP 的行为进行了研究,该技术能够实现限制、温度变化和统计分析。可以在 ATPS 中观察到 ELP 的空间组织。此外,温度变化触发了 ELP 丰富的液滴在 ATPS 中动态形成和分布,这是由于构象的变化。蛋白质与 ELP 一起被包封在合成的原细胞中,并观察到这些蛋白质和 ELP 在 ATPS 中的不同分配特性。因此,ELP 随温度凝聚的能力可以在细胞模拟系统中保持。