Journal article
Drug interactions with Bacillus anthracis topoisomerase IV: biochemical basis for quinolone action and resistance
Biochemistry (Easton), Vol.51(1), pp.370-381
01/10/2012
DOI: 10.1021/bi2013905
PMCID: PMC3261753
PMID: 22126453
Abstract
Bacillus anthracis, the causative agent of anthrax, is considered a serious threat as a bioweapon. The drugs most commonly used to treat anthrax are quinolones, which act by increasing the levels of DNA cleavage mediated by topoisomerase IV and gyrase. Quinolone resistance most often is associated with specific serine mutations in these enzymes. Therefore, to determine the basis for quinolone action and resistance, we characterized wild-type B. anthracis topoisomerase IV, the GrlA(S81F) and GrlA(S81Y) quinolone-resistant mutants, and the effects of quinolones and a related quinazolinedione on these enzymes. Ser81 is believed to anchor a water-Mg(2+) bridge that coordinates quinolones to the enzyme through the C3/C4 keto acid. Consistent with this hypothesized bridge, ciprofloxacin required increased Mg(2+) concentrations to support DNA cleavage by GrlA(S81F) topoisomerase IV. The three enzymes displayed similar catalytic activities in the absence of drugs. However, the resistance mutations decreased the affinity of topoisomerase IV for ciprofloxacin and other quinolones, diminished quinolone-induced inhibition of DNA religation, and reduced the stability of the enzyme-quinolone-DNA ternary complex. Wild-type DNA cleavage levels were generated by mutant enzymes at high quinolone concentrations, suggesting that increased drug potency could overcome resistance. 8-Methyl-quinazoline-2,4-dione, which lacks the quinolone keto acid (and presumably does not require the water-Mg(2+) bridge to mediate protein interactions), was more potent than quinolones against wild-type topoisomerase IV and was equally efficacious. Moreover, it maintained high potency and efficacy against the mutant enzymes, effectively inhibited DNA religation, and formed stable ternary complexes. Our findings provide an underlying biochemical basis for the ability of quinazolinediones to overcome clinically relevant quinolone resistance mutations in bacterial type II topoisomerases.
Details
- Title: Subtitle
- Drug interactions with Bacillus anthracis topoisomerase IV: biochemical basis for quinolone action and resistance
- Creators
- Katie J Aldred - Vanderbilt UniversitySylvia A McPhersonPengfei WangRobert J KernsDavid E GravesCharles L Turnbough JrNeil Osheroff
- Resource Type
- Journal article
- Publication Details
- Biochemistry (Easton), Vol.51(1), pp.370-381
- DOI
- 10.1021/bi2013905
- PMID
- 22126453
- PMCID
- PMC3261753
- ISSN
- 0006-2960
- eISSN
- 1520-4995
- Grant note
- R01 GM033944-27 / NIGMS NIH HHS T32 CA09582 / NCI NIH HHS R01 AI087671 / NIAID NIH HHS R56 AI081775 / NIAID NIH HHS R01 GM033944 / NIGMS NIH HHS AI81775 / NIAID NIH HHS R01 GM033944-28 / NIGMS NIH HHS T32 CA009582 / NCI NIH HHS GM33944 / NIGMS NIH HHS AI87671 / NIAID NIH HHS
- Language
- English
- Date published
- 01/10/2012
- Academic Unit
- Pharmaceutical Sciences and Experimental Therapeutics; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984366026602771
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