Journal of Biological Chemistry
Volume 278, Issue 46, 14 November 2003, Pages 45661-45671
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Enzyme Catalysis and Regulation
Identification, Characterization, and Inhibition of Plasmodium falciparum β-Hydroxyacyl-Acyl Carrier Protein Dehydratase (FabZ)*

https://doi.org/10.1074/jbc.M304283200Get rights and content
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The emergence of drug-resistant forms of Plasmodium falciparum emphasizes the need to develop new antimalarials. In this context, the fatty acid biosynthesis (FAS) pathway of the malarial parasite has recently received a lot of attention. Due to differences in the fatty acid biosynthesis systems of Plasmodium and man, this pathway is a good target for the development of new and selective therapeutic drugs directed against malaria. In continuation of these efforts we report cloning and overexpression of P. falciparum β-hydroxyacyl-acyl carrier protein (ACP) dehydratase (PffabZ) gene that codes for a 17-kDa protein. The enzyme catalyzes the dehydration of β-hydroxyacyl-ACP to trans-2-acyl-ACP, the third step in the elongation phase of the FAS cycle. It has a K m of 199 μM and k cat/K m of 80.4 m-1 s-1 for the substrate analog β-hydroxybutyryl-CoA but utilizes crotonoyl-CoA, the product of the reaction, more efficiently (K m = 86 μM, k cat/K m = 220 m-1 s-1). More importantly, we also identify inhibitors (NAS-91 and NAS-21) for the enzyme. Both the inhibitors prevented the binding of crotonoyl-CoA to PfFabZ in a competitive fashion. Indeed these inhibitors compromised the growth of P. falciparum in cultures and inhibited the parasite fatty acid synthesis pathway both in cell-free extracts as well as in situ. We modeled the structure of PfFabZ using Escherichia coli β-hydroxydecanoyl thioester dehydratase (EcFabA) as a template. We also modeled the inhibitor complexes of PfFabZ to elucidate the mode of binding of these compounds to FabZ. The discovery of the inhibitors of FabZ, reported for the first time against any member of this family of enzymes, essential to the type II FAS pathway opens up new avenues for treating a number of infectious diseases including malaria.

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The nucleotide sequence(s) reported in this paper has been submitted to the GenBank/EBI Data Bank with accession number(s) AY118082.

*

This work was supported by a grant from the Department of Biotechnology, Government of India (to N. S.) and in part by Shantha Biotechnics Pvt. Ltd., Hyderabad, India. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

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Both authors made equal contributions to this work.