A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite

Sayaka Nakashima, Zhe Liu, Yuya Yamaguchi, Shunya Saiki, Shintaro Munemasa, Toshiyuki Nakamura, Yoshiyuki Murata, Yoshimasa Nakamura

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

3,4-Dihydroxyphenylacetic acid (DOPAC) is one of the colonic microflora-produced catabolites of quercetin 4'-glucoside (Q4'G). Although the interaction of DOPAC with cellular proteins might be involved in its biological activity, the actual proteins have not yet been identified. In this study, we developed a novel tag-free DOPAC probe to label the targeted proteins by the copper(I)-catalyzed azide alkyne cycloaddition (CuAAC) and verified its efficacy. Various labeled proteins were detected by the DOPAC probe with the azide labeled biotin and a horseradish peroxidase (HRP)-streptavidin complex. Furthermore, a pulldown assay identified Keap1 and aryl hydrocarbon receptor (AhR) as the target proteins for the phase 2 enzyme up-regulation.

Original languageEnglish
Pages (from-to)240-245
Number of pages6
JournalBiochemistry and Biophysics Reports
Volume7
DOIs
Publication statusPublished - Sep 1 2016

Fingerprint

3,4-Dihydroxyphenylacetic Acid
Flavonoids
Molecules
Azides
Proteins
Aryl Hydrocarbon Receptors
Alkynes
Streptavidin
Cycloaddition
Cycloaddition Reaction
Horseradish Peroxidase
Biotin
Bioactivity
Labels
Copper
Assays
Up-Regulation
Enzymes

Keywords

  • 3,4-dihydroxyphenylacetic acid (DOPAC)
  • Click chemistry
  • Keap1
  • Quercetin
  • Tag free probe

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite. / Nakashima, Sayaka; Liu, Zhe; Yamaguchi, Yuya; Saiki, Shunya; Munemasa, Shintaro; Nakamura, Toshiyuki; Murata, Yoshiyuki; Nakamura, Yoshimasa.

In: Biochemistry and Biophysics Reports, Vol. 7, 01.09.2016, p. 240-245.

Research output: Contribution to journalArticle

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AU - Munemasa, Shintaro

AU - Nakamura, Toshiyuki

AU - Murata, Yoshiyuki

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