Detecting the Interaction of Peptide Ligands with Plant Membrane Receptors

Sarah Refi Hind, Jason S. Hoki, Joshua A. Baccile, Patrick C. Boyle, Frank C. Schroeder, Gregory B. Martin

Research output: Contribution to journalArticlepeer-review

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 enables addition of the reporter molecule after receptor-ligand binding which reduces the need for bulky ligand modifications that could interfere with the interaction.

Original languageEnglish (US)
Pages (from-to)240-269
Number of pages30
JournalCurrent protocols in plant biology
Volume2
Issue number3
DOIs
StatePublished - Sep 1 2017
Externally publishedYes

Keywords

  • click chemistry
  • ligand
  • photo-crosslinking
  • plant receptor

ASJC Scopus subject areas

  • Medicine(all)

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