Identification and functional study of a novel 2-cys peroxiredoxin (BmTPx-1) of Babesia microti

Exp Parasitol. 2016 Nov:170:21-27. doi: 10.1016/j.exppara.2016.08.005. Epub 2016 Aug 24.

Abstract

Babesia microti is an emerging human pathogen and the primary causative agent of human babesiosis in many regions of the world. Although the peroxiredoxins (Prxs) or thioredoxin peroxidases (TPx) enzymes of this parasite have been sequenced and annotated, their biological properties remain largely unknown. Prxs are a family of antioxidant enzymes that protect biological molecules against metabolically produced reactive oxygen species (ROS) and reduce hydrogen peroxide (H2O2) to water in both eukaryotes and prokaryotes. In this study, TPx-1 cDNA was cloned from B. microti (designated BmTPx-1). Recombinant BmTPx-1 (rBmTPx-1) was expressed in Escherichia coli as a histidine fusion protein and purified using Ni-NTA His bind resin. To test the defense capacity of enzymatic antioxidants against the effect of ROS, a mixed-function oxidation system was utilized with the recombinant BmTPx-1 protein. A decreased ability of rBmTPx-1 to donate electrons to the thioredoxin (Trx)/TrxR reductase system was clarified by reaction with H2O2. These results suggest that BmTPx-1 has a great impact on protecting parasites from oxidative stress in the erythrocytic stage.

Keywords: Antioxidant enzyme; Babesia microti; Peroxiredoxin.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antioxidants / chemistry
  • Antioxidants / isolation & purification*
  • Antioxidants / metabolism
  • Babesia microti / classification
  • Babesia microti / enzymology*
  • Babesia microti / genetics
  • Base Sequence
  • Blotting, Western
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescent Antibody Technique
  • Hydrogen Peroxide / metabolism
  • Mice
  • Peroxiredoxins / chemistry
  • Peroxiredoxins / genetics
  • Peroxiredoxins / isolation & purification*
  • Peroxiredoxins / metabolism
  • Phylogeny
  • Reactive Oxygen Species / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism

Substances

  • Antioxidants
  • DNA, Complementary
  • Reactive Oxygen Species
  • Recombinant Proteins
  • Hydrogen Peroxide
  • Peroxiredoxins