Carnrot, Cecilia
- Department of Animal Biosciences, Swedish University of Agricultural Sciences
Research article2008Peer reviewedOpen access
Carnrot, Cecilia; Wang, Liya; Topalis, Dimitri; Eriksson, Staffan
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 IC(50) 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.
Bacillus anthracis; thymidylate kinase; TMPK; structure model; nucleoside analogs; drug design; FMAU
Protein Science
2008, Volume: 17, number: 9, pages: 1486-1493
Publisher: COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
SDG3 Ensure healthy lives and promote well-being for all at all ages
Animal and Dairy Science
Veterinary Science
DOI: https://doi.org/10.1110/ps.034199.107
https://res.slu.se/id/publ/20799