Potential anti-infective targets in pathogenic yeasts: structure and properties of 3,4-dihydroxy-2-butanone 4-phosphate synthase of Candida albicans

J Mol Biol. 2004 Aug 20;341(4):1085-96. doi: 10.1016/j.jmb.2004.06.053.

Abstract

A synthetic gene specifying a putative 3,4-dihydroxy-2-butanone 4-phosphate synthase of Candida albicans directed the synthesis of a 22.5 kDa peptide in a recombinant Escherichia coli strain. The recombinant protein was purified to apparent homogeneity by two chromatographic steps and was shown to catalyze the formation of L-3,4-dihydroxy-2-butanone 4-phosphate from ribulose 5-phosphate at a rate of 332 nmol mg(-1) min(-1). Hydrodynamic studies indicated a native molecular mass of 41 kDa in line with a homodimer structure. The protein was crystallized in its apoform. Soaking yielded crystals in complex with the substrate ribulose 5-phosphate. The structures were solved at resolutions of 1.6 and 1.7 angstroms, respectively, using 3,4-dihydroxy-2-butanone 4-phosphate synthase of E. coli for molecular replacement. Structural comparison with the orthologs of Magnaporthe grisea and Methanococcus jannaschii revealed a hitherto unknown conformation of the essential acidic active-site loop.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antifungal Agents / pharmacology*
  • Base Sequence
  • Candida albicans / enzymology*
  • Cloning, Molecular
  • DNA, Fungal
  • Models, Molecular
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Peptide Synthases / chemistry
  • Peptide Synthases / drug effects
  • Peptide Synthases / genetics
  • Peptide Synthases / metabolism*
  • Sequence Homology, Amino Acid

Substances

  • Antifungal Agents
  • DNA, Fungal
  • 3,4-dihydroxy-2-butanone 4-phosphate synthase, C albicans
  • Peptide Synthases

Associated data

  • PDB/1TKS
  • PDB/1TKU