Crystal structures of Toxoplasma gondii pterin-4a-carbinolamine dehydratase and comparisons with mammalian and parasite orthologues

Scott Cameron, Stewart A. Fyffe, Simon Goldie, William N. Hunter

    Research output: Contribution to journalArticle

    5 Citations (Scopus)

    Abstract

    The enzyme pterin4a-carbinolamine dehydratase (PCD) is important for the recycling of pterins within eukaryotic cells. A recombinant expression system for PCD from the apicomplexan parasite Toxoplasma gondii has been prepared, the protein purified and crystallised. Single crystal X-ray diffraction methods have produced a high-resolution structure (1.6 angstrom) of the apo-enzyme and a low-resolution structure (3.1 angstrom) of a complex with a substrate-like ligand dihydrobiopterin (BH2). Analysis of the hydrogen bonding interactions that contribute to binding BH2 suggest that the ligand is present in an enol tautomeric state, which makes it more similar to the physiological substrate. The enzyme can process (R)- and (S)-forms of pterin-4a-carbinolamine and the ligand complex suggests that His61 and His79 are placed to act independently as general bases for catalysis of the individual enantiomers. Comparisons with orthologues from other protozoan parasites (Plasmodium falciparum and Leishmania major) and with rat PCD, for which the structure is known, indicate a high degree of sequence and structure conservation of this enzyme. The molecular determinants of ligand recognition and PCD reactivity are therefore highly conserved across species. (c) 2007 Elsevier B.V. All rights reserved.

    Original languageEnglish
    Pages (from-to)131-138
    Number of pages8
    JournalMolecular and Biochemical Parasitology
    Volume158
    Issue number2
    DOIs
    Publication statusPublished - Apr 2008

    Keywords

    • apicomplexa
    • crystal structure
    • dehydratase
    • kinetoplastida
    • pterin
    • tautomer
    • Toxoplasma
    • TRANSCRIPTIONAL COACTIVATOR
    • REDUCTASE
    • DCOH
    • IDENTIFICATION
    • REFINEMENT
    • MECHANISM
    • ENZYME

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