uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
Mechanisms of substrate selectivity for Bacillus anthracis thymidylate kinase
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology.
2008 (English)In: Protein Science, ISSN 0961-8368, Vol. 17, no 9, 1486-1493 p.Article in journal (Refereed) Published
Abstract [en]

Bacillus anthracis is well known in connection with biological warfare. The search for new drug targets and antibiotics is highly motivated because of upcoming multiresistant strains. Thymidylate kinase is an ideal target since this enzyme is at the junction of the de novo and salvage synthesis of dTTP, an essential precursor for DNA synthesis. Here the expression and characterization of thymidylate kinase from B. anthracis (Ba-TMPK) is presented. The enzyme phosphorylated deoxythymidine-5'-monophosphate (dTMP) efficiently with K-m and V-max values of 33 mu M and 48 mu mol mg(-1) min(-1), respectively. The efficiency of deoxyuridine-5'-monophosphate phosphorylation was; similar to 10% of that of dTMP. Several dTMP analogs were tested, and D-FMAUMP (2'-fluoroarabinosyl-5-methyldeoxyuridine-5'- monophosphate) was selectively phosphorylated with an efficiency of 172% of that of D-dTMP, but L-FMAUMP was a poor substrate as were 5-fluorodeoxyuridine-5'-monophosphate (5FdUMP) and 2',3'-dideoxy-2',3'-didehydrothymidine-5'-monophosphate (d4TMP). No activity could be detected with 3'-azidothymidine-5'-monophosphate (AZTMP). The corresponding nucleosides known as efficient anticancer and antiviral compounds were also tested, and D-FMAU was a strong inhibitor with an IC50 value of 10 mu M, while other nucleosides-L-FMAU, dThd, 5-FdUrd, d4T, and AZT, and 2'-arabinosylthymidine-were poor inhibitors. A structure model was built for Ba-TMPK based on the Staphylococcus aureus TMPK structure. Docking with various substrates suggested mechanisms explaining the differences in substrate selectivity of the human and the bacterial TMPKs. These results may serve as a start point for development of new antibacterial agents.

Place, publisher, year, edition, pages
2008. Vol. 17, no 9, 1486-1493 p.
Keyword [en]
Bacillus anthracis, thymidylate kinase, TMPK, structure model, nucleoside analogs, drug design, FMAU
National Category
Biological Sciences
URN: urn:nbn:se:uu:diva-149349DOI: 10.1110/ps.034199.107ISI: 000258651900003PubMedID: 18523102OAI: oai:DiVA.org:uu-149349DiVA: diva2:404698
Available from: 2011-03-18 Created: 2011-03-18 Last updated: 2011-03-18Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMed
By organisation
Department of Cell and Molecular Biology
In the same journal
Protein Science
Biological Sciences

Search outside of DiVA

GoogleGoogle Scholar

Altmetric score

Total: 143 hits
ReferencesLink to record
Permanent link

Direct link