Molecular and Functional Characterization of Thioredoxin 1 from 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) encoding a 107 aa protein. The deduced RuTrx amino acid sequence indicated a characteristic redox active site, (31)WCGPC(35). 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.
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea. tks1010@hanmail.net.
Julan
J
Kim
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea. moonjy@pknu.ac.kr.
Ji Young
JY
Moon
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea. wj2464@korea.kr.
Woo-Jin
WJ
Kim
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea. combikola@korea.kr.
Dong-Gyun
DG
Kim
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea. nambohye@korea.kr.
Bo-Hye
BH
Nam
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea. yobest12@korea.kr.
Young-Ok
YO
Kim
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea. genome@korea.kr.
Jung Youn
JY
Park
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea. ancm@korea.kr.
Cheul Min
CM
An
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea. heejkong@korea.kr.
Hee Jeong
HJ
Kong
D000595:Amino Acid Sequence; D000818:Animals; D000975:Antioxidants / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D003530:Cyprinidae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D053837:DNA Cleavage; D029941:Fish Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D010802:Phylogeny; D017382:Reactive Oxygen Species / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013879:Thioredoxins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
D000975:Antioxidants; D029941:Fish Proteins; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D013879:Thioredoxins
Korean rose bitterling; MCO assay; ROS detection; Rhodues uyekii; anti-oxidant; expression analysis; thioredoxin