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Thioredoxin domain-containing protein 12 from Oplegnathus fasciatus: Molecular characterization, expression against immune stimuli, and biological activities related to oxidative stress

Thulasitha, William Shanthakumar, Kim, Yucheol, Umasuthan, Navaneethaiyer, Jayasooriya, R.G.P.T., Kim, Gi-Young, Nam, Bo-Hye, Noh, Jae Koo, Lee, Jehee
Fish & shellfish immunology 2016 v.54 pp. 11-21
Edwardsiella tarda, Oplegnathus fasciatus, Rock bream iridovirus, Streptococcus iniae, active sites, amino acids, antioxidant activity, antioxidants, bream, cell viability, exons, genes, hydrogen peroxide, immune response, insulin, introns, lipopolysaccharides, liver, messenger RNA, oxidative stress, recombinant fusion proteins, transcription (genetics)
Thioredoxin domain-containing protein 12 (TXNDC12) is a small, disulfide-containing protein that belongs to the thioredoxin (TXN) superfamily. In the present study, we identified and characterized a TXNDC12-like gene, designated OfTXNDC12, from rock bream, Oplegnathus fasciatus. OfTXNDC12 consists of seven exons interrupted by six introns. Comparative genomic structural analysis revealed that the TXNDC12 of vertebrates is a structurally conserved gene. The coding sequence of OfTXNDC12 comprises 522 bp, which encodes 173 amino acid residues with the conserved thioredoxin active site motif, CGAC, and a probable C-terminal ER retrieval motif, GDEL. Transcriptional analysis of OfTXNDC12 showed the highest concentrations of the mRNA transcript in the liver, implying that it has a significant role in the liver under normal physiological conditions. In comparison, injection of lipopolysaccharide, Edwardsiella tarda, Streptococcus iniae, polyinosinic:polycytidylic acid (poly[I:C]) and rock bream iridovirus mostly triggered greater upregulation of OfTXNDC12 transcript levels in liver than in gill tissue, supporting its potential functional importance in the liver. Insulin disulfide reduction assay showed that the recombinant fusion protein (rOfTXNDC12) possesses significant thioredoxin activity. Treatment of LNCaP cells with the recombinant protein along with H2O2 revealed that rOfTXNDC12 increased the viability of cells and further supported its putative antioxidant capacity. Taken together, the results from our study suggest that OfTXNDC12 encodes for a potent antioxidant involved in redox regulation that shows significant responses to immune stimuli.