Evidence for the presence of prostaglandin H synthase like enzyme in female Setaria cervi and its inhibition by diethylcarbamazine.
Experimental evidence has shown that Setaria cervi a bovine filarial parasite contains significant amount of prostaglandin H synthase like activity in the somatic extract of its different life stages. A protein with characteristics of prostaglandin H synthase was purified to homogeneity from female somatic extract using a combination of affinity and gel filtration chromatography. Molecular weight of purified enzyme was 70kDa as determined by SDS-PAGE. Purified enzyme showed high activity with arachidonic acid and TMPD substrates suggests the presence of both cyclooxygenase and peroxidase activity in enzyme. Fluorescence spectroscopy and hemin-associated peroxidase activity confirmed presence of heme in purified enzyme. The K(m) and V(max) values using arachidonic acid were determined to be 79+/-1.5microM and 0.165+/-0.2U/ml, respectively. Further, indomethacin and aspirin, specific inhibitors for PGHS, significantly inhibited the enzyme activity. Diethylcarbamazine, an antifilarial drug inhibited the microfilarial PGHS like activity as well as their motility. Here we are reporting for the first time PGHS like activity in filarial parasite and its inhibition with DEC which provide that this enzyme could be used as a drug target.
Rathaur
Sushma
S
Department of Biochemistry, Faculty of Science, Banaras Hindu University, UP, India. surathaur@rediffmail.com
Singh
Alka
A
Yadav
Marshleen
M
Rai
Reeta
R
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D002417:Cattle; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003067:Coenzymes / Q000032:analysis; D004049:Diethylcarbamazine / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D005369:Filaricides / Q000494:pharmacology*; D005370:Filarioidea / Q000187:drug effects* / Q000201:enzymology*; D015801:Helminth Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006418:Heme / Q000032:analysis; D007700:Kinetics; D008297:Male; D011451:Prostaglandin-Endoperoxide Synthases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013772:Tetramethylphenylenediamine / Q000378:metabolism
D003067:Coenzymes; D004791:Enzyme Inhibitors; D005369:Filaricides; D015801:Helminth Proteins; D016718:Arachidonic Acid; D006418:Heme; D011451:Prostaglandin-Endoperoxide Synthases; D013772:Tetramethylphenylenediamine; D004049:Diethylcarbamazine