Abstract The ability to detect the interaction partners of a protein and how those interactions occur are central to the study of biology. Indeed, such knowledge can provide important clues to unravel the functional context of proteins in specific pathways. This review provides an overview of the different techniques available to plant researchers to study protein–protein interactions (PPIs), and briefly summarizes key benefits and disadvantages of each technique. We discuss the most commonly used in vitro-based techniques (i.e., co-immunoprecipitation and microscale thermophoresis), yeast-based screening methods (i.e., yeast two-hybrid and the split-ubiquitin system), as well as imaging and FRET-based approaches (i.e., bimolecular fluorescence complementation, modification of intracellular localisation, protein mobility measurements, acceptor photobleaching, fluorescence lifetime imaging microscopy, and fluorescence polarization/anisotropy) that are used in the field of plant biology. Finally, we offer suggestions that may provide future directions for how to best use and combine these techniques.

中文翻译：

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研究植物蛋白质-蛋白质相互作用的工具箱

摘要 检测蛋白质相互作用伙伴以及这些相互作用如何发生的能力是生物学研究的核心。事实上，这些知识可以提供重要的线索来解开特定途径中蛋白质的功能背景。本综述概述了植物研究人员可用于研究蛋白质-蛋白质相互作用 (PPI) 的不同技术，并简要总结了每种技术的主要优点和缺点。我们讨论了最常用的基于体外的技术（即免疫共沉淀和微量热泳）、基于酵母的筛选方法（即酵母双杂交和分裂泛素系统），以及基于成像和 FRET 的技术。方法（即双分子荧光互补、细胞内定位的修改、蛋白质迁移率测量、受体光漂白、荧光寿命成像显微镜和荧光偏振/各向异性），用于植物生物学领域。最后，我们提供的建议可能为如何最好地使用和组合这些技术提供未来方向。