植物パターン認識受容体のリン酸化およびタンパク質相互作用の免疫沈降法による同定

Plasma membrane-localized pattern-recognition receptors (PRRs) play a crucial role in detecting pathogen-associated molecular patterns (PAMPs). Among them, leucine-rich repeat (LRR) receptor-like kinases (RLKs) are particularly important in plants. However, their structural complexity and membrane association pose significant challenges for biochemical analysis. Many LRR-RLKs exhibit low expression levels in planta, and their hydrophobic surfaces contribute to instability, often requiring detergent-based extraction. These properties complicate co-immunoprecipitation (Co-IP), hindering the identification of interacting proteins and post-translational modifications. Recent advancements in immunoprecipitation technologies have improved the isolation of membrane proteins, enabling the analysis of protein-protein interactions and phosphorylation dynamics. Here, we describe an optimized immunoprecipitation protocol for studying plant plasma membrane receptor-like kinases, facilitating robust characterization of their interactomes and regulatory modifications.