Abstract
The bacterial system for fatty acid biosynthesis (FAS) contains several enzymes whose sequence and structure are highly conserved across a vast array of pathogens. This, coupled with their low homology and difference in organization compared to the equivalent system in humans, makes the FAS pathway an excellent target for antimicrobial drug development. To this end, we have cloned, expressed, and purified the β-hydroxyacyl-acyl carrier protein dehydratase (FabZ) from both Francisella tularensis (FtFabZ) and Yersinia pestis (YpFabZ). We also solved the crystal structures and performed an enzymatic characterization of both enzymes and several mutant forms of YpFabZ. Additionally, we have discovered two novel inhibitors of FabZ, mangostin and stictic acid, which show similar potencies against both YpFabZ and FtFabZ. Lastly, we selected several compounds from the literature that have been shown to be active against single homologues of FabZ and tested them against both YpFabZ and FtFabZ. These results have revealed clues as to which scaffolds are likely to lead to broad-spectrum antimicrobials targeted against FabZ as well as modifications to existing FabZ inhibitors that may improve potency.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Amino Acid Substitution
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Bacterial Proteins / antagonists & inhibitors
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Bacterial Proteins / chemistry*
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism
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Biocatalysis
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Catalytic Domain
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Crystallography, X-Ray
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Dimerization
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Enzyme Inhibitors / chemistry
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Enzyme Inhibitors / pharmacology
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Francisella tularensis / enzymology*
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Heterocyclic Compounds, 4 or More Rings / chemistry
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Heterocyclic Compounds, 4 or More Rings / pharmacology
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Histidine / chemistry
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Hydro-Lyases / antagonists & inhibitors
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Hydro-Lyases / chemistry*
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Hydro-Lyases / genetics
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Hydro-Lyases / metabolism
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Hydrophobic and Hydrophilic Interactions
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Models, Molecular*
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Molecular Docking Simulation
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Molecular Weight
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Oxepins / chemistry
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Oxepins / pharmacology
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Point Mutation
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Protein Conformation
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Xanthones / chemistry
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Xanthones / pharmacology
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Yersinia pestis / enzymology*
Substances
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Bacterial Proteins
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Enzyme Inhibitors
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Heterocyclic Compounds, 4 or More Rings
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Oxepins
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Recombinant Proteins
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Xanthones
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stictic acid
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Histidine
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Hydro-Lyases
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beta-hydroxyacyl-(acyl-carrier-protein)dehydrase
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mangostin