Molecular characterization and functional analysis of a peroxiredoxin 1 cDNA from golden pompano (Trachinotus ovatus)

被引：26

作者：

Wang, Long ^{[1
,2
]}

Guo, Huayang ^{[1
]}

Zhang, Nan ^{[1
]}

Ma, Zhenhua ^{[1
]}

Jiang, Shigui ^{[1
]}

Zhang, Dianchang ^{[1
]}

机构：

[1] Chinese Acad Fishery Sci, South China Sea Fisheries Res Inst, Minist Agr, Key Lab South China Sea Fishery Resources Exploit, Guangzhou 510300, Guangdong, Peoples R China

[2] Shanghai Ocean Univ, Coll Fisheries & Life Sci, Shanghai 201306, Peoples R China

来源：

DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY | 2015年 / 51卷 / 02期

关键词：

Peroxiredoxin; Trachinotus ovatus; mRNA expression; Recombinant protein; Antioxidant function; EXPRESSION ANALYSIS; REACTIVE OXYGEN; 2-CYS PEROXIREDOXIN; OXIDATIVE STRESS; IMMUNE-RESPONSE; CLONING; GENE; MECHANISMS; INNATE; THIOREDOXIN;

D O I：

10.1016/j.dci.2015.03.011

中图分类号：

S9 [水产、渔业];

学科分类号：

0908 ;

摘要：

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 I 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. (C) 2015 Elsevier Ltd. All rights reserved.

引用

页码：261 / 270

页数：10

共 49 条

[1] The oligomeric conformation of peroxiredoxins links redox state to function [J].