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 language | English (US) |
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Pages (from-to) | 240-269 |
Number of pages | 30 |
Journal | Current protocols in plant biology |
Volume | 2 |
Issue number | 3 |
DOIs | |
State | Published - Sep 1 2017 |
Externally published | Yes |
Keywords
- click chemistry
- ligand
- photo-crosslinking
- plant receptor
ASJC Scopus subject areas
- General Medicine