DXP Synthase-Catalyzed CN Bond Formation: Nitroso Substrate Specificity Studies Guide Selective Inhibitor Design
| Autor: | Francine Morris, Jürgen Bosch, Ryan J. Vierling, Caren L. Freel Meyers, Lauren E. Boucher |
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| Rok vydání: | 2013 |
| Předmět: |
Nitrogen
Stereochemistry Biochemistry Article Substrate Specificity chemistry.chemical_compound Biosynthesis Transferases Catalytic Domain Enzyme Inhibitors Molecular Biology chemistry.chemical_classification ATP synthase biology Terpenes Organic Chemistry Mutagenesis Active site Nitroso Pyruvate dehydrogenase complex Carbon Kinetics Enzyme chemistry Biocatalysis biology.protein Molecular Medicine DXP synthase |
| Zdroj: | ChemBioChem. 14:1309-1315 |
| ISSN: | 1439-4227 |
| Popis: | 1-Deoxy-D-xylulose 5-phosphate (DXP) synthase catalyzes the first step in the nonmammalian isoprenoid biosynthetic pathway to form DXP from pyruvate and D-glyceraldehyde 3-phosphate (D-GAP) in a thiamin diphosphate-dependent manner. Its unique structure and mechanism distinguish DXP synthase from its homologues and suggest that it should be pursued as an anti-infective drug target. However, few reports describe any development of selective inhibitors of this enzyme. Here, we reveal that DXP synthase catalyzes C-N bond formation and exploit aromatic nitroso substrates as active site probes. Substrate specificity studies reveal a high affinity of DXP synthase for aromatic nitroso substrates compared to the related ThDP-dependent enzyme pyruvate dehydrogenase (PDH). Results from inhibition and mutagenesis studies indicate that nitroso substrates bind to E. coli DXP synthase in a manner distinct from that of D-GAP. Our results suggest that the incorporation of aryl acceptor substrate mimics into unnatural bisubstrate analogues will impart selectivity to DXP synthase inhibitors. As a proof of concept, we show selective inhibition of DXP synthase by benzylacetylphosphonate (BnAP). |
| Databáze: | OpenAIRE |
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