Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase

Jaldappagari Seetharamappa, Muse Oke, Huanting Liu, Stephen A McMahon, Kenneth A Johnson, Lester Carter, Mark Dorward, Michal Zawadzki, Ian M Overton, C A Johannes van Niekirk, Shirley Graham, Catherine H Botting, Garry L Taylor, Malcolm F White, Geoffrey J Barton, Peter J Coote, James H Naismith

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    4 Citations (Scopus)

    Abstract

    Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase with a molecular weight of 48,168 Da, was overexpressed in methicillin-resistant Staphylococcus aureus compared with a methicillin-sensitive strain. The protein was expressed in Escherichia coli, purified and crystallized. The protein crystallized in a primitive orthorhombic Laue group with unit-cell parameters a = 83.6, b = 91.3, c = 106.0 A, alpha = beta = gamma = 90 degrees. Analysis of the systematic absences along the three principal axes indicated the space group to be P2(1)2(1)2(1). A complete data set was collected to 2.5 A resolution.

    Original languageEnglish
    Pages (from-to)452-6
    Number of pages5
    JournalActa Crystallographica F-Structural Biology and Crystallization Communications
    Volume63
    Issue numberPt 5
    DOIs
    Publication statusPublished - 1 May 2007

    Keywords

    • Bacterial Proteins
    • Chromatography, Affinity
    • Cloning, Molecular
    • Crystallization
    • Methicillin Resistance
    • Protein Conformation
    • Staphylococcus aureus
    • Transaminases

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    Seetharamappa, J., Oke, M., Liu, H., McMahon, S. A., Johnson, K. A., Carter, L., Dorward, M., Zawadzki, M., Overton, I. M., van Niekirk, C. A. J., Graham, S., Botting, C. H., Taylor, G. L., White, M. F., Barton, G. J., Coote, P. J., & Naismith, J. H. (2007). Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase. Acta Crystallographica F-Structural Biology and Crystallization Communications, 63(Pt 5), 452-6. https://doi.org/10.1107/S1744309107019562