Cloning, production and characterisation of a recombinant Cu/Zn superoxide dismutase from Taenia solium

Alejandro Castellanos-González, Lucía Jiménez, Abraham Landa

Research output: Contribution to journalArticlepeer-review

23 Scopus citations

Abstract

A full-length complementary DNA clone encoding a cytosolic Cu/Zn superoxide dismutase with a Mr of 15,588 Da was isolated from a Taenia solium larvae complementary DNA library. Comparison analysis of its deduced amino acid sequence revealed a 71% identity with Schistosoma mansoni, 57.2-59.8% with mammalian and less than 54% with other helminth cytosolic Cu/Zn superoxide dismutase. The characteristic motifs and the amino acid residues involved in coordinating copper and zinc enzymatic function are conserved. The T. solium Cu/Zn superoxide dismutase was expressed in the pRSET vector. Enzymatic and filtration chromatographic analysis showed a recombinant enzyme with an activity of 2,941 U/mg protein and a native Mr of 37 kDa. Inhibition assays using KCN, H2O2, NaN3 and SDS indicated that Cu/Zn is the metallic cofactor in the enzyme. Thiabendazole (500 μM) and albendazole (300 μM) completely inhibited the activity of T. solium Cu/Zn superoxide dismutase. Thiabendazole had no effect on bovine Cu/Zn superoxide dismutase; in contrast, albendazole had a moderate effect on it at same concentrations. Antibodies against T. solium Cu/Zn superoxide dismutase did not affect the enzymatic function; nevertheless, it cross reacts with several Taenia species, but not with trematodes, nematodes, pig, human and bovine Cu/Zn superoxide dismutase enzymes. Western blot analysis indicated the enzyme was expressed in all stages. These results indicate that T. solium possesses a Cu/Zn superoxide dismutase enzyme that can protect him from oxidant-damage caused by the superoxide anion.

Original languageEnglish (US)
Pages (from-to)1175-1182
Number of pages8
JournalInternational Journal for Parasitology
Volume32
Issue number9
DOIs
StatePublished - 2002

Keywords

  • Albendazole
  • Cestoda
  • Complementary DNA
  • Superoxide dismutase
  • Taenia solium
  • Thiabendazole

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

Fingerprint Dive into the research topics of 'Cloning, production and characterisation of a recombinant Cu/Zn superoxide dismutase from Taenia solium'. Together they form a unique fingerprint.

Cite this