Isolation of a nitric oxide synthase from the protozoan parasite, Leishmania donovani.
A soluble nitric oxide synthase (NOS) activity was purified 2800-fold from Leishmania donovani, the causative parasite of visceral leishmaniasis, by two-step affinity and anion-exchange chromatography. The purified enzyme ran as a prominent band of 110 kDa on SDS-PAGE whereas gel filtration experiments estimated the native molecular mass to be 230 +/- 20 kDa indicating that the native enzyme exists as a dimer. The enzyme activity required NADPH and was blocked by EGTA. The enzyme kinetics, cofactor requirements, inhibition studies and Western blot analysis with brain anti-NOS antibody suggest its similarity with mammalian NOS isoform I.
Basu
N K
NK
Molecular Cell Biology Division, Indian Institute of Chemical Biology, Calcutta, India.
Kole
L
L
Ghosh
A
A
Das
P K
PK
D000818:Animals; D019281:Dimerization; D004533:Egtazic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D007893:Leishmania donovani / Q000201:enzymology*; D008322:Mammals; D008970:Molecular Weight; D009249:NADP / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D004791:Enzyme Inhibitors; D007527:Isoenzymes; D004533:Egtazic Acid; D009249:NADP; D019001:Nitric Oxide Synthase