Spatiotemporal control of coacervate formation within liposomes
Spatiotemporal control of coacervate formation within liposomes
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DOI：
10.1038/s41467-019-09855-x
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发表时间：
2019-04-17
影响因子：
16.6
通讯作者：
Dekker, Cees
中科院分区：
文献类型：
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作者：
Deshpande, Siddharth;Brandenburg, Frank;Dekker, Cees
液-液相分离（LLPS），特别是凝聚，在细胞生物学中起着至关重要的作用，因为它在细胞中形成许多无膜细胞器。凝聚体在调节细胞内生物化学中起着不可或缺的作用，其功能障碍与多种疾病有关。LLPS动力学的理解将极大地受益于模拟细胞的受控体外测定。在这里，我们使用基于微流体的方法来形成凝聚体内细胞大小（类似于10 μ m）的脂质体，允许控制动力学。蛋白质孔介导的小分子向脂质体中的渗透被动地或经由如核酸的酶促聚合的主动机制触发LLPS。我们证明了蛋白质（FtsZ）和超分子组装（脂质囊泡）的螯合，以及凝聚体内宿主代谢反应（β-半乳糖苷酶活性）的可能性。这种脂质体中的凝聚物平台提供了一种了解细胞内相行为的通用工具，这些混合系统将允许工程复杂的途径来重建细胞功能，并促进自下而上创建合成细胞。
Liquid-liquid phase separation (LLPS), especially coacervation, plays a crucial role in cell biology, as it forms numerous membraneless organelles in cells. Coacervates play an indispensable role in regulating intracellular biochemistry, and their dysfunction is associated with several diseases. Understanding of the LLPS dynamics would greatly benefit from controlled in vitro assays that mimic cells. Here, we use a microfluidics-based methodology to form coacervates inside cell-sized (similar to 10 mu m) liposomes, allowing control over the dynamics. Protein-pore-mediated permeation of small molecules into liposomes triggers LLPS passively or via active mechanisms like enzymatic polymerization of nucleic acids. We demonstrate sequestration of proteins (FtsZ) and supramolecular assemblies (lipid vesicles), as well as the possibility to host metabolic reactions (beta-galactosidase activity) inside coacervates. This coacervate-in-liposome platform provides a versatile tool to understand intracellular phase behavior, and these hybrid systems will allow engineering complex pathways to reconstitute cellular functions and facilitate bottom-up creation of synthetic cells.