Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Campylobacter jejuni

J.A. Musso-Buendía, A.E. Vidal, J. Carrero-Lérida, L.M. Ruiz-Pérez, D. González-Pacanowska, G. Kasinthan, C. Nguyen, I.H. Gilbert, N.G. Johansson, Keith Wilson

    Research output: Contribution to journalArticle

    5 Citations (Scopus)

    Abstract

    The enzyme deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase) catalyses the hydrolysis of dUTP to dUMP and PPi thus controlling the incorporation of uracil into DNA genomes. In Campylobacter jejuni dUTPase exhibits structural properties of dimeric proteins characteristic of protozoa of the Kinetoplastidae family. In the present study we perform a kinetic analysis of Campylobacter dUTPase using the continuous spectrophotometric method and show that the enzyme is highly specific for deoxyuridine nucleotides. The Michaelis-Menten constant for dUTP was 0.66 µM while the k was 12.3 s. dUDP was also efficiently hydrolysed although the specificity constant, k/K, was five fold lower than for dUTP. The reaction product and the non hydrolysable analogue a,ß imido dUDP are potent inhibitors of the enzyme while several analogues of dUMP with substituents at the 3'- and 5'-positions active against trimeric dUTPases, show poor inhibitory activity. Apparent structural and kinetic differences with other eukaryotic dUTPases suggest that the present enzyme might be exploited as a target for new drugs against campylobacteriosis.
    Original languageEnglish
    Pages (from-to)111-116
    Number of pages6
    JournalJournal of Enzyme Inhibition and Medicinal Chemistry
    Volume24
    Issue number1
    DOIs
    Publication statusPublished - 1 Jan 2009

    Fingerprint

    Campylobacter jejuni
    Enzymes
    Deoxyuridine
    Campylobacter
    Uracil
    Enzyme Inhibitors
    Hydrolysis
    Nucleotides
    Genome
    DNA
    Pharmaceutical Preparations
    deoxyuridine triphosphate
    Proteins
    2'-deoxyuridylic acid

    Cite this

    Musso-Buendía, J. A., Vidal, A. E., Carrero-Lérida, J., Ruiz-Pérez, L. M., González-Pacanowska, D., Kasinthan, G., ... Wilson, K. (2009). Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Campylobacter jejuni. Journal of Enzyme Inhibition and Medicinal Chemistry, 24(1), 111-116. https://doi.org/10.1080/14756360801915476
    Musso-Buendía, J.A. ; Vidal, A.E. ; Carrero-Lérida, J. ; Ruiz-Pérez, L.M. ; González-Pacanowska, D. ; Kasinthan, G. ; Nguyen, C. ; Gilbert, I.H. ; Johansson, N.G. ; Wilson, Keith. / Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Campylobacter jejuni. In: Journal of Enzyme Inhibition and Medicinal Chemistry. 2009 ; Vol. 24, No. 1. pp. 111-116.
    @article{6e785547a6834bd09095c58f521b934b,
    title = "Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Campylobacter jejuni",
    abstract = "The enzyme deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase) catalyses the hydrolysis of dUTP to dUMP and PPi thus controlling the incorporation of uracil into DNA genomes. In Campylobacter jejuni dUTPase exhibits structural properties of dimeric proteins characteristic of protozoa of the Kinetoplastidae family. In the present study we perform a kinetic analysis of Campylobacter dUTPase using the continuous spectrophotometric method and show that the enzyme is highly specific for deoxyuridine nucleotides. The Michaelis-Menten constant for dUTP was 0.66 µM while the k was 12.3 s. dUDP was also efficiently hydrolysed although the specificity constant, k/K, was five fold lower than for dUTP. The reaction product and the non hydrolysable analogue a,{\ss} imido dUDP are potent inhibitors of the enzyme while several analogues of dUMP with substituents at the 3'- and 5'-positions active against trimeric dUTPases, show poor inhibitory activity. Apparent structural and kinetic differences with other eukaryotic dUTPases suggest that the present enzyme might be exploited as a target for new drugs against campylobacteriosis.",
    author = "J.A. Musso-Buend{\'i}a and A.E. Vidal and J. Carrero-L{\'e}rida and L.M. Ruiz-P{\'e}rez and D. Gonz{\'a}lez-Pacanowska and G. Kasinthan and C. Nguyen and I.H. Gilbert and N.G. Johansson and Keith Wilson",
    note = "MEDLINE{\circledR} is the source for the MeSH terms of this document.",
    year = "2009",
    month = "1",
    day = "1",
    doi = "10.1080/14756360801915476",
    language = "English",
    volume = "24",
    pages = "111--116",
    journal = "Journal of Enzyme Inhibition and Medicinal Chemistry",
    issn = "1475-6366",
    publisher = "Informa Healthcare",
    number = "1",

    }

    Musso-Buendía, JA, Vidal, AE, Carrero-Lérida, J, Ruiz-Pérez, LM, González-Pacanowska, D, Kasinthan, G, Nguyen, C, Gilbert, IH, Johansson, NG & Wilson, K 2009, 'Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Campylobacter jejuni', Journal of Enzyme Inhibition and Medicinal Chemistry, vol. 24, no. 1, pp. 111-116. https://doi.org/10.1080/14756360801915476

    Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Campylobacter jejuni. / Musso-Buendía, J.A.; Vidal, A.E.; Carrero-Lérida, J.; Ruiz-Pérez, L.M.; González-Pacanowska, D.; Kasinthan, G.; Nguyen, C.; Gilbert, I.H.; Johansson, N.G.; Wilson, Keith.

    In: Journal of Enzyme Inhibition and Medicinal Chemistry, Vol. 24, No. 1, 01.01.2009, p. 111-116.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Campylobacter jejuni

    AU - Musso-Buendía, J.A.

    AU - Vidal, A.E.

    AU - Carrero-Lérida, J.

    AU - Ruiz-Pérez, L.M.

    AU - González-Pacanowska, D.

    AU - Kasinthan, G.

    AU - Nguyen, C.

    AU - Gilbert, I.H.

    AU - Johansson, N.G.

    AU - Wilson, Keith

    N1 - MEDLINE® is the source for the MeSH terms of this document.

    PY - 2009/1/1

    Y1 - 2009/1/1

    N2 - The enzyme deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase) catalyses the hydrolysis of dUTP to dUMP and PPi thus controlling the incorporation of uracil into DNA genomes. In Campylobacter jejuni dUTPase exhibits structural properties of dimeric proteins characteristic of protozoa of the Kinetoplastidae family. In the present study we perform a kinetic analysis of Campylobacter dUTPase using the continuous spectrophotometric method and show that the enzyme is highly specific for deoxyuridine nucleotides. The Michaelis-Menten constant for dUTP was 0.66 µM while the k was 12.3 s. dUDP was also efficiently hydrolysed although the specificity constant, k/K, was five fold lower than for dUTP. The reaction product and the non hydrolysable analogue a,ß imido dUDP are potent inhibitors of the enzyme while several analogues of dUMP with substituents at the 3'- and 5'-positions active against trimeric dUTPases, show poor inhibitory activity. Apparent structural and kinetic differences with other eukaryotic dUTPases suggest that the present enzyme might be exploited as a target for new drugs against campylobacteriosis.

    AB - The enzyme deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase) catalyses the hydrolysis of dUTP to dUMP and PPi thus controlling the incorporation of uracil into DNA genomes. In Campylobacter jejuni dUTPase exhibits structural properties of dimeric proteins characteristic of protozoa of the Kinetoplastidae family. In the present study we perform a kinetic analysis of Campylobacter dUTPase using the continuous spectrophotometric method and show that the enzyme is highly specific for deoxyuridine nucleotides. The Michaelis-Menten constant for dUTP was 0.66 µM while the k was 12.3 s. dUDP was also efficiently hydrolysed although the specificity constant, k/K, was five fold lower than for dUTP. The reaction product and the non hydrolysable analogue a,ß imido dUDP are potent inhibitors of the enzyme while several analogues of dUMP with substituents at the 3'- and 5'-positions active against trimeric dUTPases, show poor inhibitory activity. Apparent structural and kinetic differences with other eukaryotic dUTPases suggest that the present enzyme might be exploited as a target for new drugs against campylobacteriosis.

    UR - http://www.scopus.com/inward/record.url?scp=58949095907&partnerID=8YFLogxK

    U2 - 10.1080/14756360801915476

    DO - 10.1080/14756360801915476

    M3 - Article

    VL - 24

    SP - 111

    EP - 116

    JO - Journal of Enzyme Inhibition and Medicinal Chemistry

    JF - Journal of Enzyme Inhibition and Medicinal Chemistry

    SN - 1475-6366

    IS - 1

    ER -

    Musso-Buendía JA, Vidal AE, Carrero-Lérida J, Ruiz-Pérez LM, González-Pacanowska D, Kasinthan G et al. Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Campylobacter jejuni. Journal of Enzyme Inhibition and Medicinal Chemistry. 2009 Jan 1;24(1):111-116. https://doi.org/10.1080/14756360801915476