Target Protein Identification on Photocatalyst-Functionalized Magnetic Affinity Beads

Michihiko Tsushima, Shinichi Sato, Keita Nakane, Hiroyuki Nakamura

Research output: Contribution to journalArticle

Abstract

Although various affinity chromatography and photoaffinity labeling methods have been developed for target protein identification of bioactive molecules, it is often difficult to detect proteins that bind the ligand with weak transient affinity using these techniques. We have developed single electron transfer–mediated tyrosine labeling using ruthenium photocatalysts. Proximity labeling using 1-methyl-4-aryl-urazole (MAUra) labels proteins in close proximity to the photocatalyst with high efficiency and selectivity. Performing this labeling reaction on affinity beads makes it possible to label proteins that bind the ligand with weak transient affinity. In this article, novel protocols are described for target protein identification using photocatalyst proximity labeling on ruthenium photocatalyst-functionalized magnetic affinity beads.

Original languageEnglish
Article numbere108
JournalCurrent Protocols in Protein Science
Volume101
Issue number1
DOIs
Publication statusPublished - 2020 Sep 1

Keywords

  • affinity chromatography
  • lectin
  • magnetic beads
  • protein chemical labeling
  • proximity labeling
  • target identification

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry

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