Zhu Ling, Huq Enamul
Department of Molecular Biosciences, The Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA.
Biological Laboratories, University of Texas at Austin, Austin, TX, USA.
Methods Mol Biol. 2019;2026:29-39. doi: 10.1007/978-1-4939-9612-4_3.
In light signaling pathways, protein complexes play essential roles in light perception and signal transduction. The phytochrome family of photoreceptors perceives red/far-red region of the light spectrum and then translocates into the nucleus to form protein complexes. Many phytochrome interacting proteins have been identified based on yeast two-hybrid screening and other protein-protein interaction methods. However, it is essential to demonstrate that these proteins interact with phytochromes in vivo to be functionally relevant. In this chapter, a protocol for demonstrating light dependent in vivo interactions between phytochromes and phytochrome interacting proteins is described. This protocol can be adapted for any putative phytochrome interacting protein for validation of their interaction in vivo.
在光信号通路中，蛋白质复合物在光感知和信号转导中发挥着重要作用。光受体的光敏色素家族感知光谱中的红/远红区域，然后转移到细胞核中形成蛋白质复合物。基于酵母双杂交筛选和其他蛋白质-蛋白质相互作用方法，已经鉴定出许多光敏色素相互作用蛋白。然而，证明这些蛋白质在体内与光敏色素相互作用具有功能相关性至关重要。在本章中，描述了一种用于证明光敏色素与光敏色素相互作用蛋白之间光依赖性体内相互作用的方法。该方法可适用于任何假定的光敏色素相互作用蛋白，以验证它们在体内的相互作用。
