Peroxiredoxins (Prxs) are cysteine-dependent antioxidant proteins that play a leading part in oxidative stress response. Peroxiredoxin 4 (Prx4) is located in the endoplasmic reticulum, where it is primarily involved in regulating the concentration of H2O2, generated during protein folding. Prx4 is also located in the extracellular space, where it activates the JAK/STAT-mediated stress response. Here, we focus on the identification and characterization of the sequence and function of Prx4 from the big-belly seahorse (Hippocampus abdominalis) (HaPrx4). The size of the HaPrx4 coding sequence was 777 bp, which encoded a 258 amino acid protein of 28.8 kDa molecular weight. The amino acid sequence comprises a signal peptide, two active sites with peroxidatic cysteine and resolving cysteine, catalytic triad, and peroxiredoxin superfamily domain. According to the tissue distribution results, ovaries exhibited the highest HaPrx4 expression level within fourteen examined tissues. The HaPrx4 transcriptional responses to four immune stimulants (lipopolysaccharides, polyinosinic: polycytidylic acid, Edwardsiella tarda, and Streptococcus iniae) were evaluated in the blood and liver tissues. Additionally, the functions of recombinant HaPrx4 protein were evaluated by metal ion-catalyzed oxidation assay, peroxidase activity assay, insulin reduction assay, cell viability assay, and Hoechst staining. The assay results confirmed that the functions of HaPrx4 involved DNA protection, hydrogen peroxide (H2O2) elimination, oxidoreductase activity, enhancing cell survival, and cell protection. The results of the current study propose that HaPrx4 is effectively involved in H2O2 scavenging activity during stress conditions and in innate immune responses of the big-belly seahorse.
Keywords: Antioxidant; Big-belly seahorse; Innate immunity; Peroxidase activity; Peroxiredoxin 4.
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