Characterisation of Fasciola hepatica cytochrome c peroxidase as an enzyme with potential antioxidant activity in vitro.
Cytochrome c peroxidase oxidises hydrogen peroxide using cytochrome c as the electron donor. This enzyme is found in yeast and bacteria and has been also described in the trematodes Fasciola hepatica and Schistosoma mansoni. Using partially purified cytochrome c peroxidase samples from Fasciola hepatica we evaluated its role as an antioxidant enzyme via the investigation of its ability to protect against oxidative damage to deoxyribose in vitro. A system containing FeIII-EDTA plus ascorbate was used to generate reactive oxygen species superoxide radical, H2O2 as well as the hydroxyl radical. Fasciola hepatica cytochrome c peroxidase effectively protected deoxyribose against oxidative damage in the presence of its substrate cytochrome c. This protection was proportional to the amount of enzyme added and occurred only in the presence of cytochrome c. Due to the low specific activity of the final partially purified sample the effects of ascorbate and calcium chloride on cytochrome c peroxidase were investigated. The activity of the partially purified enzyme was found to increase between 10 and 37% upon reduction with ascorbate. However, incubation of the partially purified enzyme with 1 mM calcium chloride did not have any effect on enzyme activity. Our results showed that Fasciola hepatica CcP can protect deoxyribose from oxidative damage in vitro by blocking the formation of the highly toxic hydroxyl radical (.OH). We suggest that the capacity of CcP to inhibit .OH-formation, by efficiently removing H2O2 from the in vitro oxidative system, may extend the biological role of CcP in response to oxidative stress in Fasciola hepatica.
Campos
E G
EG
Institute of Parasitology, McGill University, Quebec, Canada. elida@unb.br
Hermes-Lima
M
M
Smith
J M
JM
Prichard
R K
RK
D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D060387:Ascorbate Peroxidases; D002122:Calcium Chloride / Q000494:pharmacology; D003574:Cytochrome c Group / Q000378:metabolism; D003578:Cytochrome-c Peroxidase / Q000187:drug effects / Q000494:pharmacology / Q000502:physiology*; D003855:Deoxyribose / Q000037:antagonists & inhibitors*; D005210:Fasciola hepatica / Q000201:enzymology*; D015801:Helminth Proteins / Q000494:pharmacology / Q000502:physiology*; D010544:Peroxidases / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D012756:Sheep; D013997:Time Factors
D000975:Antioxidants; D003574:Cytochrome c Group; D015801:Helminth Proteins; D017382:Reactive Oxygen Species; D003855:Deoxyribose; D010544:Peroxidases; D060387:Ascorbate Peroxidases; D003578:Cytochrome-c Peroxidase; D002122:Calcium Chloride