Molecular and Functional Characterization of Thioredoxin 1from Korean Rose Bitterling (Rhodeus uyekii)
Thioredoxin is a multifunctional antioxidant enzyme that belongs to the reductase family. In this study, we cloned and characterized thioredoxin 1 cDNA from the Korean rose bitterling Rhodeus uyekii (RuTrx). The full-length RuTrx cDNA consists of 674 bp with a 324 nt open reading frame (ORF) encodin...
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MDPI AG
2015-08-01
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| Series: | International Journal of Molecular Sciences |
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| author | Julan Kim Ji Young Moon Woo-Jin Kim Dong-Gyun Kim Bo-Hye Nam Young-Ok Kim Jung Youn Park Cheul Min An Hee Jeong Kong |
| author_facet | Julan Kim Ji Young Moon Woo-Jin Kim Dong-Gyun Kim Bo-Hye Nam Young-Ok Kim Jung Youn Park Cheul Min An Hee Jeong Kong |
| author_sort | Julan Kim |
| collection | DOAJ |
| description | Thioredoxin is a multifunctional antioxidant enzyme that belongs to the reductase family. In this study, we cloned and characterized thioredoxin 1 cDNA from the Korean rose bitterling Rhodeus uyekii (RuTrx). The full-length RuTrx cDNA consists of 674 bp with a 324 nt open reading frame (ORF) encoding a 107 aa protein. The deduced RuTrx amino acid sequence indicated a characteristic redox active site, 31WCGPC35. Pairwise alignment revealed RuTrx amino acid identity (55.1%–83.2%) with orthologs from various species of mammalia, amphibia, fish and bird. Phylogenetic analysis was conducted to determine the evolutionary position of RuTrx. Expression analysis showed that RuTrx transcripts were present in all of the tissues examined, and was high in the hepatopancreas of R. uyekii. During early development, the expression of RuTrx transcripts was increased. Recombinant RuTrx protein (rRuTrx) was tested for its capacity to serve as an antioxidant enzyme using a metal-catalyzed oxidation (MCO) system. The ability of rRuTrx to protect against supercoiled DNA cleavage due to oxidative nicking increased in a dose-dependent manner. In Raw264.7 cells, Dihydroethidium (DHE) staining for ROS production indicated the antioxidant activity of rRuTrx. Together, these findings suggest that RuTrx may play a role in maintaining the redox state balance in Korean rose bitterling R. uyekii. |
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| spelling | doaj.art-a7fb81f25760480aa53b0b76f6e6e6042022-12-22T02:46:53ZengMDPI AGInternational Journal of Molecular Sciences1422-00672015-08-01168194331944610.3390/ijms160819433ijms160819433Molecular and Functional Characterization of Thioredoxin 1from Korean Rose Bitterling (Rhodeus uyekii)Julan Kim0Ji Young Moon1Woo-Jin Kim2Dong-Gyun Kim3Bo-Hye Nam4Young-Ok Kim5Jung Youn Park6Cheul Min An7Hee Jeong Kong8Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, KoreaBiotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, KoreaBiotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, KoreaBiotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, KoreaBiotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, KoreaBiotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, KoreaBiotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, KoreaBiotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, KoreaBiotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, KoreaThioredoxin is a multifunctional antioxidant enzyme that belongs to the reductase family. In this study, we cloned and characterized thioredoxin 1 cDNA from the Korean rose bitterling Rhodeus uyekii (RuTrx). The full-length RuTrx cDNA consists of 674 bp with a 324 nt open reading frame (ORF) encoding a 107 aa protein. The deduced RuTrx amino acid sequence indicated a characteristic redox active site, 31WCGPC35. Pairwise alignment revealed RuTrx amino acid identity (55.1%–83.2%) with orthologs from various species of mammalia, amphibia, fish and bird. Phylogenetic analysis was conducted to determine the evolutionary position of RuTrx. Expression analysis showed that RuTrx transcripts were present in all of the tissues examined, and was high in the hepatopancreas of R. uyekii. During early development, the expression of RuTrx transcripts was increased. Recombinant RuTrx protein (rRuTrx) was tested for its capacity to serve as an antioxidant enzyme using a metal-catalyzed oxidation (MCO) system. The ability of rRuTrx to protect against supercoiled DNA cleavage due to oxidative nicking increased in a dose-dependent manner. In Raw264.7 cells, Dihydroethidium (DHE) staining for ROS production indicated the antioxidant activity of rRuTrx. Together, these findings suggest that RuTrx may play a role in maintaining the redox state balance in Korean rose bitterling R. uyekii.http://www.mdpi.com/1422-0067/16/8/19433thioredoxinKorean rose bitterling Rhodues uyekiiexpression analysisanti-oxidantMCO assayROS detection |
| spellingShingle | Julan Kim Ji Young Moon Woo-Jin Kim Dong-Gyun Kim Bo-Hye Nam Young-Ok Kim Jung Youn Park Cheul Min An Hee Jeong Kong Molecular and Functional Characterization of Thioredoxin 1from Korean Rose Bitterling (Rhodeus uyekii) International Journal of Molecular Sciences thioredoxin Korean rose bitterling Rhodues uyekii expression analysis anti-oxidant MCO assay ROS detection |
| title | Molecular and Functional Characterization of Thioredoxin 1from Korean Rose Bitterling (Rhodeus uyekii) |
| title_full | Molecular and Functional Characterization of Thioredoxin 1from Korean Rose Bitterling (Rhodeus uyekii) |
| title_fullStr | Molecular and Functional Characterization of Thioredoxin 1from Korean Rose Bitterling (Rhodeus uyekii) |
| title_full_unstemmed | Molecular and Functional Characterization of Thioredoxin 1from Korean Rose Bitterling (Rhodeus uyekii) |
| title_short | Molecular and Functional Characterization of Thioredoxin 1from Korean Rose Bitterling (Rhodeus uyekii) |
| title_sort | molecular and functional characterization of thioredoxin 1from korean rose bitterling rhodeus uyekii |
| topic | thioredoxin Korean rose bitterling Rhodues uyekii expression analysis anti-oxidant MCO assay ROS detection |
| url | http://www.mdpi.com/1422-0067/16/8/19433 |
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