DXP synthase-catalyzed c-n bond formation

Nitroso substrate specificity studies guide selective inhibitor design

Francine Morris, Ryan Vierling, Lauren Boucher, Jürgen Bosch, Caren L Meyers

Research output: Contribution to journalArticle

Abstract

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).

Original languageEnglish (US)
Pages (from-to)1309-1315
Number of pages7
JournalChemBioChem
Volume14
Issue number11
DOIs
StatePublished - Jul 2013

Fingerprint

Substrate Specificity
Substrates
Glyceraldehyde 3-Phosphate
Pyruvic Acid
Thiamine Pyrophosphate
Mutagenesis
Biosynthetic Pathways
Terpenes
Enzyme Inhibitors
Study Guide
deoxyxylulose-5-phosphate synthase
Escherichia coli
Catalytic Domain
Oxidoreductases
Enzymes
Pharmaceutical Preparations

Keywords

  • Biosynthesis
  • DXP synthase
  • Enzyme inhibitors
  • Isoprenoids
  • Kinetics
  • Substrate specificity

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Molecular Medicine
  • Molecular Biology

Cite this

DXP synthase-catalyzed c-n bond formation : Nitroso substrate specificity studies guide selective inhibitor design. / Morris, Francine; Vierling, Ryan; Boucher, Lauren; Bosch, Jürgen; Meyers, Caren L.

In: ChemBioChem, Vol. 14, No. 11, 07.2013, p. 1309-1315.

Research output: Contribution to journalArticle

Morris, Francine ; Vierling, Ryan ; Boucher, Lauren ; Bosch, Jürgen ; Meyers, Caren L. / DXP synthase-catalyzed c-n bond formation : Nitroso substrate specificity studies guide selective inhibitor design. In: ChemBioChem. 2013 ; Vol. 14, No. 11. pp. 1309-1315.
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