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Detecting the Interaction of Peptide Ligands with Plant Membrane Receptors

Detecting the Interaction of Peptide Ligands with Plant Membrane Receptors 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. © 2017 by John Wiley & Sons, Inc. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Current Protocols in Plant Biology Wiley

Detecting the Interaction of Peptide Ligands with Plant Membrane Receptors

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References (38)

Publisher
Wiley
Copyright
© John Wiley and Sons
ISSN
2379-8068
eISSN
2379-8068
DOI
10.1002/cppb.20053
Publisher site
See Article on Publisher Site

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. © 2017 by John Wiley & Sons, Inc.

Journal

Current Protocols in Plant BiologyWiley

Published: Sep 1, 2017

Keywords: ; ; ;

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