title: Molecular and biochemical characterisation of ornithine decarboxylases in the sheep abomasal nematode parasites Teladorsagia circumcincta and Haemonchus contortus.
abstract: Full length cDNA encoding ornithine decarboxylases (ODC; EC 4.1.1.17) were cloned from the sheep abomasal nematode parasites Teladorsagia circumcincta (TcODC) and Haemonchus contortus (HcODC). The TcODC (1272 bp) and HcODC cDNA (1266 bp) encoded 424 and 422 amino acid proteins respectively. The predicted TcODC amino acid sequence showed 87% identity with HcODC and 65% and 64% with Caenorhabditis elegans and Caenorhabditis briggsae ODC respectively. All binding sites and active regions were completely conserved in both proteins. Soluble N-terminal His-tagged ODC proteins were expressed in Escherichia coli strain BL21, purified and characterised. The recombinant TcODC and HcODC had very similar kinetic properties: K(m) ornithine was 0.2-0.25 mM, optimum [PLP] was 0.3 mM and the pH optima were pH 8. No enzyme activity was detected when arginine was used as substrate. One millimolar difluoromethylornithine (DFMO) completely inhibited TcODC and HcODC activity, whereas 2 mM agmatine did not inhibit activity. The present study showed that ODC is a separate enzyme from arginine decarboxylase and strictly uses ornithine as substrate.
authors.affiliation: AgResearch Ltd, Hopkirk Research Institute, Grasslands Research Centre, Private Bag 11-008, Palmerston North 4442, New Zealand. saleh.umair@agresearch.co.nz
authors.forename: Saleh
authors.initials: S
authors.lastname: Umair
authors.forename: Jacqueline S
authors.initials: JS
authors.lastname: Knight
authors.forename: Heather V
authors.initials: HV
authors.lastname: Simpson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D006189:Haemonchus / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D009955:Ornithine Decarboxylase / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D012756:Sheep / Q000469:parasitology*; D014251:Trichostrongyloidea / Q000201:enzymology* / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D009955:Ornithine Decarboxylase