Purification and characterization of 3-hydroxymethylglutaryl-coenzyme A reductase of Schistosoma mansoni: regulation of parasite enzyme activity differs from mammalian host.
The enzyme 3-hydroxymethylglutaryl-CoA (HMG-CoA) reductase plays a critical role in regulating the production of cholesterol, dolichols, and ubiquinones in mammals. The inhibition of this enzyme in Schistosoma mansoni is accompanied by a cessation of egg production by the female parasite and a reduced ability of the parasite to properly glycoslyate their proteins. Furthermore, we recently demonstrated that mevinolin, if given continuously over a period of 10-14 days, is a potent antischistosomal drug. In this paper, we describe the properties of purified HMG-CoA reductase from S. mansoni. Using affinity chromatography, we were able to obtain a 417-fold purification of the enzyme which had Km values similar to the rat enzyme for HMG-CoA and NADPH. The Ki value for mevinolin, a potent and selective inhibitor of the rat reductase (Ki = 0.6 nM), was significantly higher (Ki = 46 nM) for the schistosome enzyme. SDS-PAGE and HPLC of the purified enzyme resulted in the appearance of a single protein, which had a molecular weight (66,000) in the range reported for the rat enzyme. Parasite reductase activity, unlike that of its host, did not display a circadian rhythm. Furthermore, agents which elevate (cholestyramine) or decrease (cholesterol) mammalian reductase activity had no effect on the parasite enzyme. Our results suggest that the mechanism which regulates production of the parasite's enzyme may differ from its mammalian host.
Chen
G Z
GZ
Department of Pharmacology and Toxicology, Michigan State University, East Lansing 48824.
Foster
L
L
Bennett
J L
JL
D000214:Acyl Coenzyme A / Q000378:metabolism; D000818:Animals; D002791:Cholesterol, Dietary / Q000008:administration & dosage; D002792:Cholestyramine Resin / Q000008:administration & dosage; D002940:Circadian Rhythm; D004789:Enzyme Activation; D005260:Female; D006903:Hydroxymethylglutaryl CoA Reductases / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008148:Lovastatin / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology; D008297:Male; D051379:Mice; D008862:Microsomes, Liver / Q000201:enzymology*; D009249:NADP / Q000378:metabolism; D051381:Rats; D012550:Schistosoma mansoni / Q000201:enzymology*; D012555:Schistosomiasis mansoni / Q000201:enzymology / Q000469:parasitology
D000214:Acyl Coenzyme A; D002791:Cholesterol, Dietary; D002792:Cholestyramine Resin; C008047:3-hydroxy-3-methylglutaryl-coenzyme A; D009249:NADP; D008148:Lovastatin; D006903:Hydroxymethylglutaryl CoA Reductases