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Cloning and functional characterization of rockfish peroxiredoxin 4 homolog with its innate immune responses.
- Source :
-
Fish & shellfish immunology [Fish Shellfish Immunol] 2019 May; Vol. 88, pp. 364-374. Date of Electronic Publication: 2019 Mar 07. - Publication Year :
- 2019
-
Abstract
- The fourth member of the typical 2-cysteine peroxiredoxin (Prx4) is a well-known antioxidant enzyme, which reduces different peroxides in their catalytic process. The present study reports the identification of the rockfish Sebastes schlegelii Prx4 (SsPrx4) at a genomic level, as well as the characterization of its structural and functional features. SsPrx4 harbors a complete ORF of 786 bp encoding a polypeptide (29 kDa) of 262 amino acids (aa) with an isoelectric point of 6.2. Thioredoxin 2 domain was prominent in the SsPrx4 sequence, which has a signal peptide (31 bp) at the N-terminus. Hence, the SsPrx4 may be functionally active in the cytoplasm of rockfish cells. Moreover, two VCP motifs and three catalytic triad residues ( <superscript>112</superscript> T, <superscript>115</superscript> C, <superscript>191</superscript> R) were identified in the SsPrx4 protein sequence. A peroxidatic cysteine ( <superscript>115</superscript> C <subscript>P</subscript> ) and resolving cysteines ( <superscript>236</superscript> C <subscript>R</subscript> ) were detected at the VCP motifs. The rockfish Prx4 genome consists of seven exons, which are similar to the architecture of other Prx4 orthologs. The deduced amino acid sequence of SsPrx4 shares a relatively high amino acid sequence identity (91.6%) and close evolutionary relationship with Miichthys miiuy and Stegastes partitus Prx4. The potential for scavenging extracellular H <subscript>2</subscript> O <subscript>2</subscript> was evidenced by the purified recombinant SsPrx4 protein (rSsPrx4) in vitro system. Moreover, rSsPrx4 may protect the plasmid DNA in a metal-catalyzed oxidation system and catalyze the reduction of an insulin disulfide bond. Quantitative real-time PCR revealed that SsPrx4 mRNA was ubiquitously expressed in fourteen different tissues, with the highest expression observed in the liver followed by the ovary, and kidney tissues. Transcriptional modulations were observed in liver and spleen tissues of rockfish after injecting them with bacterial stimuli, including Streptococcus iniae, LPS, and a viral mimic of poly I:C. Together, the results suggest that SsPrx4 may play an important role in both the antioxidant and innate immune defense of black rockfish. These findings provide structural and functional insights into the SsPrx4 of the teleost.<br /> (Copyright © 2019. Published by Elsevier Ltd.)
- Subjects :
- Animals
Antioxidants metabolism
Cloning, Molecular
Female
Fish Proteins genetics
Hydrogen Peroxide
Kidney metabolism
Liver metabolism
Male
Ovary metabolism
Perciformes genetics
Peroxiredoxins genetics
Phylogeny
Sequence Alignment
Streptococcal Infections immunology
Streptococcus iniae
Fish Proteins immunology
Immunity, Innate
Perciformes immunology
Peroxiredoxins immunology
Streptococcal Infections veterinary
Subjects
Details
- Language :
- English
- ISSN :
- 1095-9947
- Volume :
- 88
- Database :
- MEDLINE
- Journal :
- Fish & shellfish immunology
- Publication Type :
- Academic Journal
- Accession number :
- 30853656
- Full Text :
- https://doi.org/10.1016/j.fsi.2019.03.010