Expression, purification and characterization of an atypical 2-Cys peroxiredoxin from the silkworm, Bombyx mori

Insect Mol Biol. 2015 Apr;24(2):203-12. doi: 10.1111/imb.12149. Epub 2014 Dec 16.

Abstract

Peroxiredoxins (Prxs) play important roles in protecting organisms against damage caused by reactive oxygen species (ROS). In this study, we cloned a cDNA of Bombyx mori peroxiredoxin 5 (BmPrx5), which contained a 565-bp open reading frame for a 188-residue protein. Sequence analysis indicated that BmPrx5 belongs to the atypical 2-Cys peroxiredoxin family. Recombinant BmPrx5 purified from Escherichia coli showed antioxidant activity that removes H2 O2 and protects DNA from oxidative damage. Quantitative real-time PCR showed that the level of BmPrx5 mRNA in haemocytes increased early and decreased by 24 h after injection of H2 O2 whereas, in the fat body, the transcript level decreased at 6 h and increased at 12 h. Pseudomonas aeruginosa and Staphylococcus aureus infection resulted in higher levels of H2 O2 in the haemolymph and of BmPrx5 mRNA in haemocytes at 8 h postinfection. These data suggest that BmPrx5 acts as an antioxidant enzyme to protect the silkworm from oxidative damage induced by bacterial infection. Further study is needed to elucidate the exact role of BmPrx5 in the silkworm immune system.

Keywords: antioxidant; immunity; peroxiredoxin; silkworm.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Bombyx / enzymology*
  • Bombyx / genetics
  • Bombyx / microbiology
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Gene Expression
  • Hemocytes / chemistry
  • Hemolymph / chemistry
  • Hydrogen Peroxide / analysis
  • Insect Proteins / genetics*
  • Insect Proteins / metabolism
  • Open Reading Frames
  • Oxidative Stress
  • Peroxiredoxins / genetics*
  • Peroxiredoxins / metabolism
  • Pseudomonas aeruginosa
  • Reactive Oxygen Species / metabolism
  • Real-Time Polymerase Chain Reaction
  • Staphylococcus aureus

Substances

  • Insect Proteins
  • Reactive Oxygen Species
  • Hydrogen Peroxide
  • Peroxiredoxins