State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, China.
Methods Mol Biol. 2021;2297:75-82. doi: 10.1007/978-1-0716-1370-2_8.
The red (R)/far-red (FR) light absorbing phytochromes are one of the major photoreceptor classes in plants. Phytochromes exist in two distinct but interconvertible forms: the R light-absorbing Pr form and the FR light-absorbing Pfr form. Upon photoactivation by light, phytochromes physically interact with their partners to transduce the light signal. Co-immunoprecipitation (Co-IP) is one of the most efficient techniques to study these protein-protein interactions in vivo. However, the co-IP procedure for phytochromes needs to be modified to allow their formation of Pr or Pfr form. Here, we describe a detailed co-IP procedure to examine which form of phytochrome (Pr or Pfr) is preferentially associated with their interacting partners in vivo.
红光/远红光吸收的光敏色素是植物中主要的光受体之一。光敏色素存在两种明显但可相互转化的形式：红光吸收的 Pr 形式和远红光吸收的 Pfr 形式。光激活后，光敏色素通过与伴侣蛋白相互作用来传递光信号。免疫共沉淀（Co-IP）是研究体内这些蛋白-蛋白相互作用最有效的技术之一。然而，光敏色素的 Co-IP 程序需要进行修改，以允许其形成 Pr 或 Pfr 形式。在这里，我们描述了一个详细的 Co-IP 程序，以检查哪种形式的光敏色素（Pr 或 Pfr）在体内与它们的相互作用伙伴优先结合。
