High-throughput screen for inhibitors of 1-deoxy-D-xylulose 5-phosphate reductoisomerase by surrogate ligand competition

Elizabeth B. Gottlin, R. Edward Benson, Scott Conary, Brett Antonio, Kellie Duke, E. Sturgis Payne, S. Salman Ashraf, Dale J. Christensen

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)


1-Deoxy-D-xylulose 5-phosphate reductoisomerase (Dxr) is a key enzyme in a biosynthetic pathway for isoprenoids that is unique to eubacteria and plants. Dxr catalyzes the rearrangement and NADPH-dependent reduction of 1-deoxy-D-xylulose 5-phosphate to 2-C-methyl-D-erythritol 4-phosphate. The authors have purified Escherichia coli Dxr and devised a high-throughput screen (HTS) for compounds that bind to this enzyme at a functional site. Evidence is presented that the surrogate ligand directly binds or allosterically affects both the D-1-deoxyxylulose 5-phosphate (DXP) and NADPH binding sites. Compounds that bind at either or both sites that compete for binding with the surrogate ligand register as hits. The time-resolved fluorescence-based assay represents an improvement over the Dxr enzyme assay that relies on relatively insensitive measurements of NADPH oxidation. Screening 32,000 compounds from a diverse historical library, the authors obtained 89 potent inhibitors in the surrogate ligand competition assay. The results presented here suggest that peptide surrogate ligands may be useful in formatting HTS for proteins with difficult biochemical assays or targets of unknown function.

Original languageEnglish
Pages (from-to)332-339
Number of pages8
JournalJournal of Biomolecular Screening
Issue number3
Publication statusPublished - Jun 2003
Externally publishedYes


  • DXP and NADPH binding sites
  • Dxr
  • Fosmidomycin
  • High-throughput screening
  • Ligand-binding assays

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Molecular Medicine
  • Pharmacology
  • Drug Discovery


Dive into the research topics of 'High-throughput screen for inhibitors of 1-deoxy-D-xylulose 5-phosphate reductoisomerase by surrogate ligand competition'. Together they form a unique fingerprint.

Cite this