A potential antioxidant enzyme belonging to the atypical 2-Cys peroxiredoxin subfamily characterized from rock bream, Oplegnathus fasciatus

• Published in: Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology Vol. 187; pp. 1 - 13
• Main Authors: Saranya Revathy, Kasthuri, Umasuthan, Navaneethaiyer, Whang, Ilson, Jung, Hyung-Bok, Lim, Bong-Soo, Nam, Bo-Hye, Lee, Jehee
• Format: Journal Article
• Language: English
• Published: England Elsevier Inc 01.09.2015
• Subjects: Amino Acid Sequence; Animals; Antioxidant protection; Antioxidants - chemistry; Antioxidants - metabolism; Base Sequence; Cloning, Molecular; DNA Damage; Escherichia coli - physiology; Evolution, Molecular; Gene Expression Regulation - drug effects; Gene structure; Genomics; Hydrogen Peroxide - pharmacology; Iridovirus - physiology; Lipopolysaccharides - pharmacology; Mitochondrial peroxiredoxin 5 (Prx5); Models, Molecular; Molecular Sequence Data; mRNA expression profiling; Oxidative stress; Perciformes - immunology; Perciformes - metabolism; Perciformes - microbiology; Perciformes - virology; Peroxiredoxins - chemistry; Peroxiredoxins - genetics; Peroxiredoxins - metabolism; Phylogeny; Poly I-C - pharmacology; Promoter Regions, Genetic - genetics; Protein Conformation; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sequence Alignment; Sequence Homology, Amino Acid; Oxidative stress; Antioxidant protection; mRNA expression profiling; Gene structure; Mitochondrial peroxiredoxin 5 (Prx5)
• ISSN: 1096-4959; 1879-1107
• DOI: 10.1016/j.cbpb.2015.04.008

Peroxiredoxins (Prxs), a diverse family of antioxidant enzymes, exert their antioxidant function through which different peroxide species are detoxified. This study describes both structural and functional characterization of a mitochondrial Prx identified in rock bream, Oplegnathus fasciatus (RbPrx5). The ORF (573 bp) of RbPrx5 encoded a protein of 190 amino acids (20kDa) containing a putative mitochondrial targeting sequence (residues 1–20) and a thioredoxin-2 motif (residues 31–190) and three conserved Cys residues. Homology assessment and phylogenetic analysis clearly disclosed relatively higher amino acid sequence similarities and a closer evolutionary position of RbPrx5 with those of other teleost homologs. The ORF of RbPrx5 was distributed among six exons as found in other vertebrates, but it possessed an additional exon in its 5'-UTR. In silico examination of RbPrx5 gene’s putative promoter region revealed the presence of several cis-elements which may be important in its transcriptional regulation. Constitutive expression of RbPrx5 was detected in eleven tissues with the highest level in the heart. Modulation of RbPrx5 transcription was evidenced from varying mRNA levels in head kidney post in vivo LPS-, poly I:C-, Edwardsiella tarda bacterial- and rock bream iridoviral-challenges. The antioxidant function of RbPrx5 was investigated using recombinant RbPrx5 protein. Results of an in vitro mixed-function oxidase assay demonstrated a dose-dependent inhibition of DNA damage by rRbPrx5. A H2O2 tolerance assay showed that in vivo overexpression of rRbPrx5 increased the bacterial survival under H2O2-mediated oxidative stress condition. These findings provide an overall insight into the structural, expressional and functional aspects of RbPrx5.