Hu Meilin, Liu Xiaoqing, Liang Taizhen, Zheng Chunfu, Ma Xiancai
Guangzhou National Laboratory, Guangzhou International Bio-Island, Guangzhou, China.
State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
Methods Mol Biol. 2025;2940:63-78. doi: 10.1007/978-1-0716-4615-1_7.
In recent years, numerous membraneless condensates have been found to regulate cellular and viral processes. These condensates are mainly composed of specific proteins with liquid-liquid phase separation (LLPS) properties. Many studies have revealed that these membraneless organelles efficiently promote viral infection, replication, assembly, and latency, demonstrating their druggable potential by targeting LLPS. In vitro LLPS assays have been widely used to characterize the biophysical properties and study the biochemical functions of phase-separated proteins. By simulating the intracellular environment, these assays can elucidate how cellular or viral proteins form dynamically reversible condensates via LLPS. Here, we describe the procedure of the in vitro LLPS assay for viral proteins, which can also be applied to study the LLPS properties of cellular proteins.
近年来，人们发现许多无膜凝聚物可调节细胞和病毒过程。这些凝聚物主要由具有液-液相分离（LLPS）特性的特定蛋白质组成。许多研究表明，这些无膜细胞器能有效促进病毒感染、复制、组装和潜伏，通过靶向LLPS显示出它们的可药物靶向潜力。体外LLPS分析已被广泛用于表征生物物理特性和研究相分离蛋白的生化功能。通过模拟细胞内环境，这些分析可以阐明细胞或病毒蛋白如何通过LLPS形成动态可逆的凝聚物。在这里，我们描述了用于病毒蛋白的体外LLPS分析方法，该方法也可用于研究细胞蛋白的LLPS特性。
