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Trypanosoma brucei thymidine kinase is a tandem protein consisting of two homologous parts, which together enable efficient substrate binding
Umeå University, Faculty of Medicine, Department of Medical Biochemistry and Biophysics. (Anders Hofer)
Umeå University, Faculty of Medicine, Department of Medical Biochemistry and Biophysics. (Anders Hofer)
Tehran University. (Reza Rofougaran)
Umeå University, Faculty of Medicine, Department of Medical Biochemistry and Biophysics. (Lars Thelander)
2012 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 287, 17628-17636 p.Article in journal, Editorial material (Refereed) Published
Abstract [en]

Trypanosoma brucei causes African sleeping sickness, a disease for which existing chemotherapies are limited by their toxicity or lack of efficacy. We have found that four parasites, including T. brucei, contain genes where two or four thymidine kinase (TK) sequences are fused into a single open reading frame. The T. brucei full-length enzyme as well as its two constituent parts, domain 1 and domain 2, were separately expressed and characterized. Of potential interest for nucleoside analog development, T. brucei TK was less discriminative against purines than human TK1 with the following order of catalytic efficiencies: thymidine > deoxyuridine ≫ deoxyinosine > deoxyguanosine. Proteins from the TK1 family are generally dimers or tetramers, and the quaternary structure is linked to substrate affinity. T. brucei TK was primarily monomeric but can be considered a two-domain pseudodimer. Independent kinetic analysis of the two domains showed that only domain 2 was active. It had a similar turnover number (k(cat)) as the full-length enzyme but could not self-dimerize efficiently and had a 5-fold reduced thymidine/deoxyuridine affinity. Domain 1, which lacks three conserved active site residues, can therefore be considered a covalently attached structural partner that enhances substrate binding to domain 2. A consequence of the non-catalytic role of domain 1 is that its active site residues are released from evolutionary pressure, which can be advantageous for developing new catalytic functions. In addition, nearly identical 89-bp sequences present in both domains suggest that the exchange of genetic material between them can further promote evolution.

Place, publisher, year, edition, pages
U.S: American Society for Biochemistry and Molecular Biology, 2012. Vol. 287, 17628-17636 p.
Keyword [en]
Tandem Thymidine kinase, Trypanosoma brucei, dTTP, Thymidine, genetic exchange
National Category
Biological Sciences
Research subject
biological chemistry
URN: urn:nbn:se:umu:diva-80895OAI: diva2:651899
Targeting the nucleotide metabolism of the mammalian pathogen, Trypanosoma brucei
Swedish Research Council
Available from: 2013-09-27 Created: 2013-09-27 Last updated: 2013-09-27Bibliographically approved

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Vodnala, MunenderRanjbarian, FarahnazThelander, Lars
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