title: Molecular characterization and functional analysis of a peroxiredoxin 1 cDNA from golden pompano (Trachinotus ovatus).
abstract: Peroxiredoxin 1 (Prx 1) is an important antioxidant protein that can protect organisms against the toxicity of reactive oxygen species. In this study, a full-length Prx 1 cDNA sequence (ToPrx 1) was identified from golden pompano (Trachinotus ovatus). The ToPrx 1 cDNA was 1049 base pairs (bp) long and contained a 5'-untranslated region (UTR) of 127 nucleotides, a 3'-UTR of 328 nucleotides, and a 594 bp open reading frame (ORF) encoding a 197 amino acid polypeptide. The ToPrx 1 protein showed strong homology (79-91%) with Prx 1 proteins from other species and contained the conserved Prx domain and the signature of the peroxidase catalytic center. Phylogenetic analysis revealed that ToPrx 1 was in the fish Prx 1 subgroup, which suggests that ToPrx 1 could belong to the 2-Cys Prx subgroup. ToPrx 1 mRNA was ubiquitously detected in all tested tissues, and its expression was comparatively high in the fin, spleen, kidney, intestine, eye, gill, and blood. The expression levels of ToPrx 1 mRNA were significantly up-regulated in liver, spleen, kidney, and intestine of golden pompano injected with Photobacterium damselae. The recombinant ToPrx 1 protein (rToPrx 1) was expressed and purified through affinity chromatography and refolded successfully using ion-exchange chromatography. The antioxidant activity assay of rToPrx 1 showed that it could reduce insulin in the presence of dithiothreitol, which suggests that the antioxidant function of rToPrx 1 is thiol dependent. This study provides useful information to help further understand the functional mechanism of Prx 1 in marine fish immunity.
authors.affiliation: Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 231 Xingang Road West, Haizhu District, Guangzhou 510300, China; College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China.
authors.forename: Long
authors.initials: L
authors.lastname: Wang
authors.affiliation: Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 231 Xingang Road West, Haizhu District, Guangzhou 510300, China.
authors.forename: Huayang
authors.initials: H
authors.lastname: Guo
authors.affiliation: Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 231 Xingang Road West, Haizhu District, Guangzhou 510300, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Zhang
authors.affiliation: Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 231 Xingang Road West, Haizhu District, Guangzhou 510300, China.
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Ma
authors.affiliation: Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 231 Xingang Road West, Haizhu District, Guangzhou 510300, China.
authors.forename: Shigui
authors.initials: S
authors.lastname: Jiang
authors.affiliation: Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 231 Xingang Road West, Haizhu District, Guangzhou 510300, China. Electronic address: zhangdch@163.com.
authors.forename: Dianchang
authors.initials: D
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004229:Dithiothreitol / Q000378:metabolism; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D016905:Gram-Negative Bacterial Infections / Q000276:immunology*; D007109:Immunity; D007328:Insulin / Q000378:metabolism; D008969:Molecular Sequence Data; D010473:Perciformes / Q000276:immunology*; D054464:Peroxiredoxins / Q000235:genetics* / Q000378:metabolism; D010776:Photobacterium / Q000276:immunology*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D000975:Antioxidants; D018076:DNA, Complementary; D029941:Fish Proteins; D007328:Insulin; D011994:Recombinant Proteins; D054464:Peroxiredoxins; D004229:Dithiothreitol
keywords: Antioxidant function; Peroxiredoxin; Recombinant protein; Trachinotus ovatus; mRNA expression