Detecting the interaction of peptide ligands with plant membrane receptors.
Curr Protoc Plant Biol
; 2: 240-269, 2017 09.
Article
en En
| MEDLINE
| ID: mdl-29098191
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.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Péptidos
/
Receptores de Péptidos
/
Química Clic
Idioma:
En
Revista:
Curr Protoc Plant Biol
Año:
2017
Tipo del documento:
Article