Rapid Identification of Peptide-Receptor-Coreceptor Complexes in Protoplasts

Secreted signaling peptides, also called peptide hormones, play crucial roles in regulating plant growth, development, and immunity. Plant peptide hormones are perceived by plasma membrane-localized receptor-like kinases (RLKs) or receptor-like proteins (RLPs) that harbor specific extracellular domains to bind and recognize the corresponding peptide ligands. Binding of a peptide ligand to its receptor usually induces the hetero-dimerization of the cognate receptor and a coreceptor, followed by the phosphorylation and activation of the receptor complex to transduce downstream signaling. Therefore, matching peptide ligands with their respective receptors/coreceptors is crucial for elucidating peptide hormone signaling pathways. In this chapter, using the RGF7 peptide-RGI4/RGI5 receptor-BAK1 coreceptor complex as an example, we describe a rapid method to identify the peptide ligand-receptor-coreceptor complexes via co-immunoprecipitation assays using recombinant proteins transiently expressed in Arabidopsis protoplasts.