Department of Chemistry, Pennsylvania State University, University Park, PA, United States.
Department of Chemistry, Pennsylvania State University, University Park, PA, United States.
Methods Enzymol. 2021;646:115-142. doi: 10.1016/bs.mie.2020.09.001. Epub 2020 Oct 14.
We discuss preparation of experimental models for multi-compartment membraneless organelles in which distinct compositions are maintained indefinitely for macromolecule-rich phases in contact with each other. These model systems are based on the physical chemistry phenomenon of complex coacervation. In complex coacervation, liquid-liquid phase separation occurs due to ion pairing interactions between oppositely charged polyelectrolytes. This mechanism can drive the associative phase separation of proteins and nucleic acids, the major macromolecular components of membraneless organelles. Here we provide examples, advice and practical considerations for the design, generation, and analysis of multi-compartment complex coacervates. These structures are of interest to compartmentalize the interior of artificial cells and as models for the intracellular membraneless organelles of biological cells.
我们讨论了多隔室无膜细胞器实验模型的制备，其中不同的组成部分在大分子丰富的相接触的情况下可以无限期地保持。这些模型系统基于复杂凝聚的物理化学现象。在复杂凝聚中，由于带相反电荷的聚电解质之间的离子配对相互作用，会发生液-液相分离。这种机制可以驱动无膜细胞器的主要大分子成分蛋白质和核酸的缔合相分离。在这里，我们提供了多隔室复杂凝聚物的设计、生成和分析的实例、建议和实际考虑因素。这些结构对于将人工细胞的内部隔室化以及作为生物细胞内无膜细胞器的模型都很有意义。
