Wang, Liya
- Department of Molecular Biosciences, Swedish University of Agricultural Sciences
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 K-m value (0.6 mu m) and a V-max of 3.3 mu mol dTMP mg(-1) min(-1), but deoxyuridine (K-m = 4.2 mu M, V-max = 4.1 mu mol 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.
Keywords
characterization; 5-fluoro-deoxyuridine growth inhibition; kinetics; thymidine
Published in
Biological Chemistry
2006, volume: 387, number: 12, pages: 1575-1581
Publisher: WALTER DE GRUYTER & CO
SLU Authors
Eriksson, Staffan
- Department of Molecular Biosciences, Swedish University of Agricultural Sciences
- Department of Molecular Biosciences, Swedish University of Agricultural Sciences
UKÄ Subject classification
Animal and Dairy Science
Veterinary Science
Publication identifier
- DOI: https://doi.org/10.1515/BC.2006.196
Permanent link to this page (URI)
https://res.slu.se/id/publ/10567