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Title of Journal: Appl Microbiol Biotechnol

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Abbravation: Applied Microbiology and Biotechnology

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Springer-Verlag

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DOI

10.1016/0020-7683(88)90044-3

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1432-0614

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A screening system for antioxidants using thioredo

Authors: S Izawa Y Inoue
Publish Date: 2003/10/31
Volume: 64, Issue: 4, Pages: 537-542
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Abstract

Previously we found that cytosolic thioredoxin is a negative regulator for an oxidative stress responsive transcription factor Yap1p yeast AP1like transcription factor ie this transcription factor is constitutively concentrated in the nucleus in the thioredoxindeficient mutant trx1Δtrx2Δ due to an impairment of the reactive oxygen speciesscavenging activity of this mutant Izawa et al 1999 J Biol Chem 27428459–28465 Based on these findings we developed a screening method to discover substances that show antioxidant activity With this method antioxidant activity was evaluated by monitoring the subcellular localization of Yap1p Since Yap1p is oxidized and accumulates in the nucleus in trx1Δtrx2Δ cells it is easy to identify antioxidant activity by observing the localization of green fluorescent protein GFPtagged Yap1p If exogenous substances taken in by trx1Δtrx2Δ cells were able to function as antioxidants to reduce the oxidized form of Yap1p GFP1Yap1p would diffuse into the cytoplasm We used this system to screen for antioxidant activity in mushrooms and found that the edible mushroom Agaricus blazei Murill is an excellent source of antioxidantsWe are grateful to Drs S Kuge Tohoku University and S Miyabe TaiAi for providing plasmids and mushrooms respectively This study was partly supported by the Kyoto University Venture Business Laboratory VBL and Biooriented Technology Research Advancement Institution BRAIN

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