Institute for Developmental Genetics, Heinrich-Heine University, Universitätsstr. 1, D-40225 Düsseldorf, Germany.
J Exp Bot. 2022 Jun 24;73(12):3866-3880. doi: 10.1093/jxb/erac139.
Molecular processes depend on the concerted and dynamic interactions of proteins, either by one-on-one interactions of the same or different proteins or by the assembly of larger protein complexes consisting of many different proteins. Here, not only the protein-protein interaction (PPI) itself, but also the localization and activity of the protein of interest (POI) within the cell is essential. Therefore, in all cell biological experiments, preserving the spatio-temporal state of one POI relative to another is key to understanding the underlying complex and dynamic regulatory mechanisms in vivo. In this review, we examine some of the applicable techniques to measure PPIs in planta as well as recent combinatorial advances of PPI methods to measure the formation of higher order complexes with an emphasis on in vivo imaging techniques. We compare the different methods and discuss their benefits and potential pitfalls to facilitate the selection of appropriate techniques by providing a comprehensive overview of how to measure in vivo PPIs in plants.
分子过程依赖于蛋白质的协同和动态相互作用,这种相互作用既可以通过相同或不同蛋白质的一对一相互作用来实现,也可以通过由许多不同蛋白质组成的更大的蛋白质复合物的组装来实现。在这里,不仅蛋白质-蛋白质相互作用(PPI)本身,而且感兴趣的蛋白质(POI)在细胞内的定位和活性都是至关重要的。因此,在所有细胞生物学实验中,相对于另一个 POI 来保存一个 POI 的时空状态是理解体内复杂和动态调节机制的关键。在这篇综述中,我们检查了一些适用于在植物体内测量 PPI 的技术,以及 PPI 方法的最新组合进展,重点介绍了体内成像技术,以测量更高阶复合物的形成。我们比较了不同的方法,并讨论了它们的优缺点,以方便通过提供如何在植物体内测量 PPI 的综合概述来选择适当的技术。