Division of Microbiology, Graduate School of Medicine, the University of Tokyo, Tokyo 113-0033, Japan.
STAR Protoc. 2021 Mar 31;2(2):100430. doi: 10.1016/j.xpro.2021.100430. eCollection 2021 Jun 18.
This protocol visualizes dynamic interaction between a transmembrane protein and an intracellular protein induced by clusterization/oligomerization of the transmembrane protein. Association-dissociation of the intracellular region of the transmembrane protein with cytoplasmic protein(s) is detected by proximity ligation assay. Since a transmembrane protein often resists extraction, biochemical analysis of its dynamic interaction with cytoplasmic effectors is cumbersome. This protocol quantitatively visualizes protein-protein interaction occurring in the membrane periphery, providing a powerful tool to elucidate signal transduction across the membrane. For complete details on the use and execution of this protocol, please refer to Ooki et al. (2019).
本方案通过可视化跨膜蛋白的簇集/寡聚化诱导的跨膜蛋白与胞内蛋白之间的动态相互作用。通过邻近连接分析检测跨膜蛋白胞内区与胞质蛋白（s）的缔合-解离。由于跨膜蛋白通常难以提取，因此其与胞质效应物的动态相互作用的生化分析较为繁琐。本方案定量可视化了发生在膜外周的蛋白-蛋白相互作用，为阐明跨膜信号转导提供了有力工具。有关本方案使用和实施的完整详细信息，请参考 Ooki 等人（2019 年）。
