A Fluorescence Anisotropy-Based Comprehensive Method for the In Vitro Screening of COI1–JAZs Agonists and Antagonists

Yousuke Takaoka, Kengo Hayashi, Kaho Suzuki, Ika Nurul Azizah, Minoru Ueda

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The phytohormone (+)-7-iso-jasmonoyl-l-isoleucine (JA-Ile) causes protein–protein interactions (PPI) between F-box Protein CORONATINE INSENSITIVE 1 (COI1) and JASMONATE ZIM DOMAIN (JAZ) transcriptional repressor. A total of 13 JAZ subtypes are encoded in the genome of Arabidopsis thaliana; however, their genetic redundancy obfuscates the individual function of each JAZ. One approach to decipher this redundant signaling network is chemical genetics, using small molecules specific to individual JAZ subtype, for which a reliable and high-throughput screening system of the ligands for all combinations of COI1–JAZs would be indispensable. In this chapter, we describe a fluorescence anisotropy-based quantitative screening system for the ligands of COI1–JAZ co-receptors. Our method is applicable to agonists and antagonists of the COI1–JAZs.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages145-160
Number of pages16
DOIs
Publication statusPublished - 2020 Jan 1

Publication series

NameMethods in Molecular Biology
Volume2085
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Affinity
  • COI1-JAZ co-receptor
  • Fluorescence anisotropy
  • JAZ degron peptide
  • Ligand screening
  • Protein–protein interaction

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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