Evaluation of Bacillus anthracis thymidine kinase as a potential target for the development of antibacterial nucleoside analogs

Cecilia Carnrot, Susan R. Vogel, Youngjoo Byun, Liya Wang, Werner Tjarks, Staffan Eriksson, Andrew J. Phipps

Research output: Contribution to journalArticlepeer-review

Abstract

Bacillus anthracis, which causes anthrax, has attracted attention because of its potential use as a biological weapon. The risk of multidrug resistance against B. anthracis increases the need for antibiotics with new molecular targets. Nucleoside analogs are well-known antiviral and anticancer prodrugs, and thymidine kinase catalyzes the rate-limiting step in the activation of pyrimidine nucleoside analogs used in chemotherapy. The thymidine kinase gene from B. anthracis Sterne strain (34F2) (Ba-TK) was cloned and expressed in E. coli, and the product was purified and characterized regarding its substrate specificity. Ba-TK phosphorylated pyrimidine nucleosides and all natural nucleoside triphosphates served as phosphate donors. Size exclusion chromatography indicated a dimeric form of Ba-TK, regardless of the presence of ATP. Thymidine was the most efficient substrate with a low Km value (0.6 μM) and a Vmax of 3.3 μmol dTMP mg-1 min -1, but deoxyuridine (Km=4.2 μM, Vmax=4.1 μmol dUMP mg-1 min-1) was also a good substrate. Several pyrimidine analogs were also tested and analogs with 5-position modifications showed higher activities compared to analogs with 3′- and N3-position modifications. Deoxyuridine analogs were the most potent inhibitors of B. anthracis growth in vitro. These results may be used to guide future development of nucleoside analogs against B. anthracis.

Original languageEnglish (US)
Pages (from-to)1575-1581
Number of pages7
JournalBiological Chemistry
Volume387
Issue number12
DOIs
StatePublished - Dec 1 2006
Externally publishedYes

Keywords

  • 5-fluoro-deoxyuridine
  • Characterization
  • Growth inhibition
  • Kinetics
  • Thymidine

ASJC Scopus subject areas

  • Biochemistry

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