Department of Bioscience and Biotechnology, Fukui Prefectural University, Fukui, Japan.
Methods Mol Biol. 2023;2652:129-143. doi: 10.1007/978-1-0716-3147-8_7.
Extracellular signals are usually perceived by membrane-localized receptors that transduce intercellular signals to activate various pathways. In plants, single transmembrane receptor kinases act as receptors for extracellular signals. Endogenous secreted peptide hormones have been recognized as novel signaling molecules, functioning through the formation of ligand-receptor pairs in plants. Recently, research on plant peptide hormone-receptor interactions based on the structural biology approach has greatly improved; however, the dissociation constant of recombinant receptor molecules expressed in insect cells using the baculovirus expression system is relatively low. We introduce here a method for creating a stable and functional homogeneous expression system for plant receptor kinases using tobacco BY-2 cells while maintaining conventional ligand-binding activity. This strategy will help improve our understanding of plant endogenous peptide ligand-receptor interactions.
细胞外信号通常被膜定位的受体感知，这些受体将细胞间信号转导，激活各种途径。在植物中，单个跨膜受体激酶作为细胞外信号的受体。内源性分泌的肽激素已被认为是新型信号分子，通过在植物中形成配体-受体对发挥作用。最近，基于结构生物学方法的植物肽激素-受体相互作用的研究取得了很大进展；然而，使用杆状病毒表达系统在昆虫细胞中表达的重组受体分子的解离常数相对较低。我们在这里介绍了一种使用烟草 BY-2 细胞创建植物受体激酶稳定且功能性均相表达系统的方法，同时保持了常规的配体结合活性。该策略将有助于我们更好地理解植物内源性肽配体-受体相互作用。
