Purification and Characterization of Nitric Oxide Synthase in Fasciola gigantica
نویسندگان
چکیده
Nitric oxide (NO) and NO-related pathways have the potential to play critical roles in the helminthes life cycle. Nitric oxide and nitric oxide synthase(NOS) in the parasitic trematode Fasciola gigantica were investigated. Two forms of F. gigantica nitric oxide synthase were detected after chromatography on CM-cellulose. The NOS II had the highest specific activity. NOS II was purified to homogeneity after acetone precipitation and Sephacryl S-200 column with 50% recovery and a 120 fold of purification. The molecular mass of NOSII was 18 kDa using Sephacryl S-200 and SDSPAGE. The Km for NOSII was 0.83 and 0.2 μM using NADPH and L-arginine respectively. NOSII + had a pH and temperature optima at 6 and 40oC respectively. The enzyme was stable up to 40 C and o lost 80% of its activity at 70 C. In addition Fe , Ba , Cu and Hg completely inhibited the o 3+ 2+ 2 + 2 + enzyme, Co , Ba and Zn cause 87, 77 and 84.7 % inhibition of NOSII activity. Ca had no effect 2+ 2+ 2+ 2+ on enzyme activity. The results indicated that the NOSII had very low resistant toward the most of metals.
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