Isolation and characterization of a thioredoxin domain-containing protein 12 from orange-spotted grouper, Epinephelus coioides

Fish Shellfish Immunol. 2012 Sep;33(3):667-73. doi: 10.1016/j.fsi.2012.06.030. Epub 2012 Jul 9.

Abstract

Thioredoxin domain-containing protein 12 (Txndc12) belongs to the thioredoxin superfamily, and has roles in redox regulation, defense against oxidative stress, refolding of disulfide-containing proteins, and regulation of transcription factors. In this study, a thioredoxin domain-containing protein 12 was cloned from the marine fish grouper, Epinephelus coioides by RACE PCR, named as Ec-Txndc12. The Ec-Txndc12 encodes 173 amino acid residues with signal peptide in its N-terminal and a thioredoxin (Trx) domain that is homologous with some genes in Mus musculus, Xenopus laveis, etc. Ec-Txndc12 mRNA is predominately expressed in liver, brain and muscle. The expression of Ec-Txndc12 was up-regulated in the liver of grouper challenged with SGIV. In order to elucidate its biological functions, Ec-Txndc12 was recombined and expressed in Escherichia coli BL21 (DE3). The rEc-Txndc12 fusion protein was demonstrated to possess the antioxidant activity. The grouper spleen (GS) cells were treated with a high concentration of rEc-Txndc12 (30 μg/ml), which significantly enhanced cells viability under oxidative damage caused by viral infection. These results together indicated that Ec-Txndc12 could function as an important antioxidant in a physiological context, and might be involved in the responses to viral challenge.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Line
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Fish Proteins / chemistry
  • Fish Proteins / genetics*
  • Fish Proteins / immunology
  • Fish Proteins / metabolism*
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Molecular Sequence Data
  • Organ Specificity
  • Perciformes / genetics*
  • Perciformes / immunology
  • Perciformes / metabolism*
  • Perciformes / virology
  • Phylogeny
  • Protein Disulfide Reductase (Glutathione) / chemistry
  • Protein Disulfide Reductase (Glutathione) / genetics*
  • Protein Disulfide Reductase (Glutathione) / immunology
  • Protein Disulfide Reductase (Glutathione) / metabolism*
  • RNA, Messenger / analysis
  • Ranavirus / physiology
  • Real-Time Polymerase Chain Reaction
  • Sequence Alignment
  • Sequence Homology, Amino Acid

Substances

  • DNA, Complementary
  • Fish Proteins
  • RNA, Messenger
  • Protein Disulfide Reductase (Glutathione)

Associated data

  • GENBANK/JQ514561