Structure of glycosomal glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma brucei determined from Laue data

Frederic MD Vellieux*, J Hajdu, Christophe LMJ Verlinde, Hillie Groendijk, RJ Read, TJ Greenhough, John W Campbel, Kor H Kalk, JA Littlechild, Herman C Warson, Wim GJ Hol

*Corresponding author for this work

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    Abstract

    The three-dimensional structure of glycosomal glyceraldehyde-3-phosphate dehydrogenase [D-glyceraldehyde-3-phosphate:NAD+ oxidoreductase (phosphorylating), EC 1.12.1.12] from the sleeping-sickness parasite Trypanosoma brucei was solved by molecular replacement at 3.2-angstrom resolution with an x-ray data set collected by the Laue method. For data collection, three crystals were exposed to the polychromatic synchrotron x-ray beam for a total of 20.5 sec. The structure was solved by using the Bacillus stearothermophilus enzyme model [Skarzynski, T., Moody, P. C. E. & Wonacott, A. J. (1987) J. Mol. Biol. 193, 171-187] with a partial data set which was 37% complete. The crystals contain six subunits per asymmetric unit, which allowed us to overcome the absence of >60% of the reflections by 6-fold density averaging. After molecular dynamics refinement, the current molecular model has an R factor of 17.6%. Comparing the structure of the trypanosome enzyme with that of the homologous human muscle enzyme, which was determined at 2.4-angstrom resolution, reveals important structural differences in the NAD binding region. These are of great interest for the design of specific inhibitors of the parasite enzyme.

    Original languageEnglish
    Pages (from-to)2355-2359
    Number of pages5
    JournalProceedings of the National Academy of Sciences of the United States of America
    Volume90
    Issue number6
    DOIs
    Publication statusPublished - 15-Mar-1993

    Keywords

    • POLYCHROMATIC X-RAY CRYSTALLOGRAPHY
    • PROTEIN CRYSTAL STRUCTURE
    • TRYPANOSOMIASIS
    • RATIONAL DRUG DESIGN
    • ALPHA-DIFLUOROMETHYLORNITHINE
    • PHOSPHATE DEHYDROGENASE
    • GLYCOLYTIC-ENZYMES
    • MOLECULAR-DYNAMICS
    • COMPUTER-GRAPHICS
    • REFINEMENT
    • RHODESIENSE
    • RESOLUTION
    • RESISTANT

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