title: Purification and enzymatic activity of an NADH-fumarate reductase and other mitochondrial activities of Leishmania parasites.
abstract: A 65 kD membrane-associated NADH-fumarate reductase subunit, which has a molecular weight similar to that of one of the enzyme subunits from bacteria, was purified from Leishmania donovani promastigotes. NADH-fumarate reductase and other mitochondrial enzymatic activities of L. major and L. donovani promastigotes and amastigotes were investigated. The presence of NADH-fumarate reductase was demonstrated in digitonin-permeabilized L. major promastigotes and mitochondria of L. major and L. donovani promastigotes and amastigotes. The activity of solubilized NADH-fumarate reductase was measured in L. major and L. donovani promastigotes. Succinate exhibited a clear concentration-dependent inhibitory effect on fumarate reductase, whereas fumarate also exhibited a clear concentration-dependent inhibitory effect on succinate dehydrogenase. The data indicate that fumarate reductase is an obligatory component of the respiratory chain of the parasite. Since the enzyme is an important component in the intermediate metabolism in the Leishmania parasite and is absent in mammalian cells, it could be a potential target for antileishmanial drugs.
authors.affiliation: Centre for Medical Parasitology, Department of Clinical Microbiology,University Hospital of Copenhagen, Rigshospitalet, Denmark. cmcmp@rh.dk
authors.forename: M
authors.initials: M
authors.lastname: Chen
authors.forename: M
authors.initials: M
authors.lastname: Bennedsen
authors.forename: L
authors.initials: L
authors.lastname: Zhai
authors.forename: A
authors.initials: A
authors.lastname: Kharazmi
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D007891:Leishmania / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology; D051379:Mice; D008928:Mitochondria / Q000201:enzymology*; D009243:NAD / Q000378:metabolism; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors
chemical_list: D009243:NAD; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; C020155:fumarate reductase (NADH); D008274:Magnesium; D002118:Calcium