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

doi:10.1016/j.bbrep.2016.06.020

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 journal › Article › peer-review

6 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 language	English
Pages (from-to)	240-245
Number of pages	6
Journal	Biochemistry and Biophysics Reports
Volume	7
DOIs	https://doi.org/10.1016/j.bbrep.2016.06.020
Publication status	Published - Sept 1 2016

Keywords

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

ASJC Scopus subject areas

Biophysics
Biochemistry

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10.1016/j.bbrep.2016.06.020

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title = "A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite",

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.",

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

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

note = "Funding Information: We thank Teruhiko Nitoda and Hiromasa Kiyota of Okayama University for their technical support and valuable discussions. We are also grateful to the SC NMR Laboratory of Okayama University and the MS Laboratory of the Faculty of Agriculture, Okayama University for assistance in NMR and MS experiments. This study was partly supported by MEXT KAKENHI Grant number 25292073 (YN) . Publisher Copyright: {\textcopyright} 2016 The Authors.",

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doi = "10.1016/j.bbrep.2016.06.020",

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T1 - A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite

AU - Nakashima, Sayaka

AU - Liu, Zhe

AU - Yamaguchi, Yuya

AU - Saiki, Shunya

AU - Munemasa, Shintaro

AU - Nakamura, Toshiyuki

AU - Murata, Yoshiyuki

AU - Nakamura, Yoshimasa

N1 - Funding Information: We thank Teruhiko Nitoda and Hiromasa Kiyota of Okayama University for their technical support and valuable discussions. We are also grateful to the SC NMR Laboratory of Okayama University and the MS Laboratory of the Faculty of Agriculture, Okayama University for assistance in NMR and MS experiments. This study was partly supported by MEXT KAKENHI Grant number 25292073 (YN) . Publisher Copyright: © 2016 The Authors.

PY - 2016/9/1

Y1 - 2016/9/1

N2 - 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.

AB - 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.

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KW - Quercetin

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

Abstract

Keywords

ASJC Scopus subject areas

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