1-Deoxy-d-xylulose 5-Phosphate Synthase Catalyzes a Novel Random Sequential Mechanism*

Background: 1-Deoxy-d-xylulose 5-phosphate (DXP) synthase is a thiamine diphosphate (ThDP)-dependent enzyme in pathogen isoprenoid biosynthesis and a potential drug target. Results: Tryptophan fluorescence and kinetic analyses show that donor and acceptor substrates bind reversibly and independently to DXP synthase. Conclusion: DXP synthase catalyzes a novel, ThDP-dependent, random sequential mechanism. Significance: Targeting the unique kinetic mechanism of DXP synthase could lead to new anti-infective agents. Emerging resistance of human pathogens to anti-infective agents make it necessary to develop new agents to treat infection. The methylerythritol phosphate pathway has been identified as an anti-infective target, as this essential isoprenoid biosynthetic pathway is widespread in human pathogens but absent in humans. The first enzyme of the pathway, 1-deoxy-d-xylulose 5-phosphate (DXP) synthase, catalyzes the formation of DXP via condensation of d-glyceraldehyde 3-phosphate (d-GAP) and pyruvate in a thiamine diphosphate-dependent manner. Structural analysis has revealed a unique domain arrangement suggesting opportunities for the selective targeting of DXP synthase; however, reports on the kinetic mechanism are conflicting. Here, we present the results of tryptophan fluorescence binding and kinetic analyses of DXP synthase and propose a new model for substrate binding and mechanism. Our results are consistent with a random sequential kinetic mechanism, which is unprecedented in this enzyme class.

• Publication year

  2011

• Venue

  Journal of Biological Chemistry

• Publication date

  2011-08-30

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1074/jbc.M111.259747PMID 21878632PMCID PMC3196101
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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