Pintor Katherine Lapis, Diepold Andreas, Angelidou Georgia, Glatter Timo
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.
Karlsruhe Institute of Technology, Institute of Applied Biosciences, Karlsruhe, Germany.
Methods Mol Biol. 2025;2953:41-57. doi: 10.1007/978-1-0716-4694-6_3.
Many critical protein-protein interactions are transient, making them challenging to study using established methods. This limitation is particularly evident in transport processes like bacterial secretion systems, where the interactions between the export machinery and the cargo proteins are inherently dynamic. In this protocol, we describe a sensitive method that allows for the identification of such interactions using the example of SycH, the chaperone of the bacterial type III secretion system (T3SS) effector YopH. Using this method in live Yersinia enterocolitica, we identified known as well as previously uncharacterized stable and transient interactors of this protein.
许多关键的蛋白质-蛋白质相互作用是短暂的，这使得使用既定方法对其进行研究具有挑战性。这种局限性在诸如细菌分泌系统等运输过程中尤为明显，在这些过程中，输出机制与货物蛋白之间的相互作用本质上是动态的。在本方案中，我们描述了一种灵敏的方法，该方法以细菌III型分泌系统（T3SS）效应器YopH的伴侣蛋白SycH为例，用于鉴定此类相互作用。在活的小肠结肠炎耶尔森菌中使用这种方法，我们鉴定出了该蛋白已知的以及先前未表征的稳定和短暂相互作用蛋白。
