Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals & College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang Province, P. R. China.
Department of Biochemistry and Chemistry, La Trobe Institute for Molecular Science, La Trobe University, Victoria, 3086, Australia.
Chem Asian J. 2023 Jul 17;18(14):e202300226. doi: 10.1002/asia.202300226. Epub 2023 May 5.
Protein-protein interactions (PPIs) play critical roles in almost all cellular signal transduction events. Characterization of PPIs without interfering with the functions of intact cells is very important for basic biology study and drug developments. However, the ability to profile PPIs especially those weak/transient interactions in their native states remains quite challenging. To this end, many endeavors are being made in developing new methods with high efficiency and strong operability. By coupling with advanced fluorescent microscopy and mass spectroscopy techniques, these strategies not only allow us to visualize the subcellular locations and monitor the functions of protein of interest (POI) in real time, but also enable the profiling and identification of potential unknown interacting partners in high-throughput manner, which greatly facilitates the elucidation of molecular mechanisms underlying numerous pathophysiological processes. In this review, we will summarize the typical methods for PPIs identification in living cells and their principles, advantages and limitations will also be discussed in detail.
蛋白质-蛋白质相互作用 (PPIs) 在几乎所有细胞信号转导事件中都起着关键作用。在不干扰完整细胞功能的情况下对 PPIs 进行表征对于基础生物学研究和药物开发非常重要。然而，在其天然状态下对 PPIs 进行分析，特别是那些弱/瞬时相互作用的能力仍然具有相当大的挑战性。为此，人们正在努力开发具有高效率和强可操作性的新方法。通过与先进的荧光显微镜和质谱技术相结合，这些策略不仅使我们能够实时可视化亚细胞位置和监测感兴趣蛋白质 (POI) 的功能，而且还能够以高通量的方式对潜在的未知相互作用伙伴进行分析和鉴定，这极大地促进了对许多病理生理过程的分子机制的阐明。在这篇综述中，我们将总结在活细胞中鉴定 PPIs 的典型方法及其原理，详细讨论它们的优点和局限性。
