Identification and localization of glutathione S-transferase as a potential target enzyme in Brugia species.
Brugia filarial nematodes are pathogenic lymphatic-dwelling parasites that, like other helminths, may modify the host's defense mechanisms by a major detoxification process involving glutathione-binding proteins such as glutathione S-transferases (GSTs). In the present study, soluble extracts of third-stage larvae, adult male and female worms, microfilariae of either B. pahangi or B. malayi or the adult worm excretory-secretory products of B. malayi were used to determine GST activity. These extracts and affinity-purified fractions of B. pahangi adult worms had a specific enzymatic activity when 1-chloro-2,4-dinitrobenzene was used as a substrate. The observance of this enzyme in all life cycle stages of Brugia spp. demonstrates its ubiquitous nature. Lavage of intraperitoneally infected jirds, but not that of uninfected jirds, also showed increased enzymatic activity, suggesting that GST is secreted in vivo. Soluble proteins of both Brugia spp. were strongly recognized by antibodies in sera from rabbits immunized with affinity-purified native GST of Onchocerca volvulus. Immunohistochemical studies localized these proteins in adult worms, demonstrating cross-reactivity between the GST of these two filarial nematodes. The effect of this enzyme on the motility and viability of adult worms, microfilariae, and larvae was tested in vitro using a battery of known GST inhibitors. Of all those tested, ethacrynic acid, N-ethylmalemide, 4-nitropyridine-oxide, or 1-chloro-2,4-dinitrobenzene at micromolar concentrations reduced the viability and motility of microfilariae, third-stage larvae, and adult worms. These results suggest that Brugia GSTs are major metabolic enzymes and may play an important role in the parasite's survival.
Rao
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UR
Department of Microbiology and Parasitology, School of Veterinary Medicine, Louisiana State University, Baton Rouge, USA.
Salinas
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Mehta
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Klei
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TR
D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology; D017178:Brugia malayi / Q000201:enzymology* / Q000254:growth & development; D017179:Brugia pahangi / Q000201:enzymology* / Q000254:growth & development; D003429:Cross Reactions; D005260:Female; D005368:Filariasis / Q000469:parasitology*; D005849:Gerbillinae; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D008018:Life Cycle Stages; D008297:Male; D017181:Onchocerca volvulus / Q000201:enzymology / Q000276:immunology
D000909:Antibodies, Helminth; D005982:Glutathione Transferase