Removal of hydrogen peroxide by a 1-cysteine peroxiredoxin enzyme of the filarial parasite Dirofilaria immitis

Prior studies have shown that filarial nematodes can effectively metabolize hydrogen peroxide in excess of that generated by activated host cells. How ever. the mechanisms of H2O2 removal by the filarial parasites are unclear. Herein we report the results of studies carried out on the biochemical act ivity and on immunolocalization of a recombinant peroxiredoxin (Prx) enzyme from the dog filarial parasite Dirofilaria immitis. A full-length cDNA enc oding a 1-Cys Prx enzyme from the dog heartworm D. immitis was expressed in Escherichia coli as a recombinant polyhistidine fusion protein (rDiPrx-1). rDiPrx-1 was capable of reducing H2O2 in the presence of dithiothreitol. T he apparent kinetic constants determined for DiPrx-1 using H2O2 as a substr ate were a Michaelis constant (K-m) of 16.28 mM and a maximal velocity (v(m ax)) of 16 mu mol min(-1). Consistent with the enzyme activity, D. immitis adult worms could detoxify exogenously added H2O2 in vitro. Antibodies to r DiPrx-1 identified a 27-kDa native antigen in parasite extracts and larval and adult excretory-secretory products. The antibodies were used to localiz e the native antigen to the lateral hypodermal chords of both male and fema le worms by immunohistochemistry. In addition, labeling was seen in the afi brillar muscle cells in male worms and in some areas of the uterine wall in female worms. Thus. DiPrx-1 is the first parasite Prx to be shown to detox ify exogenously added H2O2 in an in vitro system.