Detecting the Interaction of Peptide Ligands with Plant Membrane Receptors

Abstract

The field of plant receptor biology has rapidly expanded in recent years, however the demonstration of direct interaction between receptor-ligand pairs remains a challenge. Click chemistry has revolutionized small molecule research but lacks popularity in plant research. Here we describe a method that tests for the direct physical interaction of a candidate receptor protein and a peptide ligand. This protocol describes the generation of the ligand probe, transient expression of a receptor protein, enrichment of membrane-bound receptors, photo-crosslinking and click chemistry-mediated reporter addition, and detection of the receptor-ligand complex. Copper-based click chemistry confers several advantages, including the versatility to use almost any azide-containing reporter molecule for detection or visualization of the complex and addition of the reporter molecule after receptor-ligand binding which reduces the need for bulky ligand modifications that could interfere with the interaction.

Non‐volatile metabolites mediate plant interactions with insect herbivores

“…In a more targeted approach, click chemistry and photo-crosslinking were successfully applied to identify the Solanum lycopersicum (tomato) receptor for flgII-28, a flagellin fragment that is characteristic of Pseudomonas syringae and other bacterial phytopathogens (Hind et al, 2016(Hind et al, , 2017. Given the widespread parallels between plant perception of herbivory-and pathogen-associated molecular patterns, we expect that similar crosslinking methods can be used identify more plant cell surface receptors perceiving insect herbivory-associated ligands.…”