Biochemical characterization of recombinant dihydroorotate dehydrogenase from the opportunistic yeast Candida albicans

Elke Zameitat, Z. Gojkovic, W. Knecht, Jure Piskur, Monica Löffler

Research output: Contribution to journalArticlepeer-review

23 Citations (SciVal)

Abstract

Candida albicans is the most prevalent yeast pathogen in humans, and recently it has become increasingly resistant to the current antifungal agents. In this study we investigated C. albicans dihydroorotate dehydrogenase (DHODH, EC 1.3.99.11), which catalyzes the fourth step of de novo pyrimidine synthesis, as a new target for controlling infection. We propose that the enzyme is a member of the DHODH family 2, which comprises mitochondrially bound enzymes, with quinone as the direct electron acceptor and oxygen as the final electron acceptor. Full-length DHODH and N-terminally truncated DHODH, which lacks the targeting sequence and the transmembrane domain, were subcloned from C. albicans, recombinantly expressed in Escherichia coli, purified, and characterized for their kinetics and substrate specificity. An inhibitor screening with 28 selected compounds was performed. Only the dianisidine derivative, redoxal, and the biphenyl quinoline-carboxylic acid derivative, brequinar sodium, which are known to be potent inhibitors of mammalian DHODH, markedly reduced C. albicans DHODH activity. This study provides a background for the development of antipyrimidines with high efficacy for decreasing in situ pyrimidine nucleotide pools in C. albicans.
Original languageEnglish
Pages (from-to)3183-3191
JournalThe FEBS Journal
Volume273
Issue number14
DOIs
Publication statusPublished - 2006

Subject classification (UKÄ)

  • Biochemistry and Molecular Biology

Keywords

  • DHODase
  • pathogenes
  • nucleic acid precursors
  • yeast

Fingerprint

Dive into the research topics of 'Biochemical characterization of recombinant dihydroorotate dehydrogenase from the opportunistic yeast Candida albicans'. Together they form a unique fingerprint.

Cite this