Identification of Carbonyl Reductase 1 as a Resveratrol-Binding Protein by Affinity Chromatography Using 4'-Amino-3,5-dihydroxy-<i>trans</i>-stilbene
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- ITO Yuta
- Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University
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- MITANI Takakazu
- Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University
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- HARADA Naoki
- Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University
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- ISAYAMA Atsushi
- Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University
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- TANIMORI Shinji
- Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University
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- TAKENAKA Shigeo
- Division of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University
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- NAKANO Yoshihisa
- Osaka Women’s Junior College
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- INUI Hiroshi
- Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University
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- YAMAJI Ryoichi
- Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University
Bibliographic Information
- Other Title
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- Identification of Carbonyl Reductase 1 as a Resveratrol-Binding Protein by Affinity Chromatography Using 4'-Amino-3,5-dihydroxy-trans-stilbene
- Identification of carbonyl reductase 1 as a resveratrol-binding protein by affinity chromatography using 4′-amino-3,5-dihydroxy-trans-stilbene
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Abstract
The mechanisms by which resveratrol (3,4',5-trihydroxy-trans-stilbene) elicits diverse health benefits remain unclear because the intracellular target molecules of resveratrol are poorly defined. We screened resveratrol-binding proteins from lysates of MCF-7 breast cancer cells using resveratrol-affinity resin, which was constructed by immobilizing 4'-amino-3,5-dihydroxy-trans-stilbene on activated CH-Sepharose. On SDS-PAGE, two bands were detected as proteins that specifically bound to the resveratrol-affinity resin. One of these, a 30-kDa protein, was identified as human carbonyl reductase 1 (CBR1) by hybrid linear ion trap/time-of-flight mass spectrometry. Similarly, recombinant CBR1 bound to the resveratrol-affinity resin in the absence of resveratrol, but not in the presence of resveratrol. Among its activities, CBR1 catalyzes a NADPH-dependent reduction of the anticancer drug doxorubicin to the cardiotoxin doxorubicinol. The effects of doxorubicin on viability of MCF-7 cells were enhanced by resveratrol, 3,5-dihydroxy-4'-methoxy-trans-stilbene, 3,4'-dihydroxy-5-methoxy-trans-stilbene, and 4'-amino-3,5-dihydroxy-trans-stilbene at concentrations of 1 and 10 μM. Resveratrol and these derivatives inhibited CBR1 activities to a similar degree at concentrations of 100 and 200 μM. However, 3,5-dimethoxy-4'-hydroxy-trans-stilbene and m-hydroquinone had no influence on doxorubicin cytotoxicity or CBR1 activity. Resveratrol inhibited CBR1 activity through an apparent mix of competitive (Ki=55.8 μM) and noncompetitive (αKi=164 μM; α=2.98) inhibition kinetics. These results indicate that (i) resveratrol enhances the cytotoxic effects of doxorubicin on MCF-7 cells; (ii) the moiety that contains the 3,5-dihydroxyl groups of resveratrol, but not the m-hydroquinone structure alone, is required to bind CBR1; and (iii) resveratrol acts as a mixed-type inhibitor of CBR1 activity on doxorubicin.
Journal
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- Journal of Nutritional Science and Vitaminology
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Journal of Nutritional Science and Vitaminology 59 (4), 358-364, 2013
Center for Academic Publications Japan
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Details 詳細情報について
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- CRID
- 1390282681301443840
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- NII Article ID
- 130004491360
- 40019766271
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- NII Book ID
- AA00703822
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- COI
- 1:CAS:528:DC%2BC3sXhsVegs7rM
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- ISSN
- 18817742
- 03014800
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- NDL BIB ID
- 024801882
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- PubMed
- 24064738
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- Abstract License Flag
- Disallowed