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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 UedaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2085)

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.

Key words

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

Notes

Acknowledgments

This work was supported by a Grant-in-Aid for Scientific Research (no. 26282207, 17H06407, 17H00885 to MU, and No. 25708026, 18H02101 to Y.T.), JSPS A3 Foresight Program (to MU), JSPS Core-to-core Program ‘Asian Chemical Biology Initiative’ (to MU), and JST PREST (JPMJPR16Q4 to YT).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Yousuke Takaoka
    • 1
    • 2
  • Kengo Hayashi
    • 1
  • Kaho Suzuki
    • 1
  • Ika Nurul Azizah
    • 1
  • Minoru Ueda
    • 1
    • 3
    Email author
  1. 1.Department of Chemistry, Graduate School of ScienceTohoku UniversitySendaiJapan
  2. 2.Precursory Research for Embryonic Science and Technology (PREST)Japan Science and Technology AgencyTokyoJapan
  3. 3.Department of Molecular and Chemical Life Sciences, Graduate School of Life SciencesTohoku UniversitySendaiJapan

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