Mechanism of inhibition of DNA gyrase by quinolone antibacterials: specificity and cooperativity of drug binding to DNA.
Although the functional target of quinolone antibacterials such as nalidixic acid and norfloxacin has been identified as the enzyme DNA gyrase, the direct binding site of the drug is the DNA molecule [Shen, L. L., & Pernet, A. G. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 307-311]. As described in this paper, binding specificity and cooperativity of quinolones to DNA were further investigated with the use of a variety of DNA species of different structures and different base compositions. Results show that the drug binding specificity is controlled and determined largely by the DNA structure. The drug binds weakly and demonstrates no base preference when DNA strands are paired. The drug binds with much greater affinity when the strands are separated, and consequently, binding preference emerges: it binds better to poly(G) and poly(dG) over their counterparts including poly(dI). The results suggest that the drug binds to unpaired bases via hydrogen bonding and not via ring stacking with DNA bases. The weak binding to relaxed double-stranded DNA and the stronger binding to single-stranded DNA are both nonspecific as they do not demonstrate binding saturation and cooperativity. The specific type of binding, initially demonstrated in our previous publication with the supercoiled DNA and more recently with complex formed between linear DNA and DNA gyrase [Shen, L. L., Kohlbrenner, W. E., Weigl, D., & Baranowski, J. (1989) J. Biol. Chem. (in press)], occurs near the drug's supercoiling inhibition concentration. As shown in this paper, binding saturation curves of this type are highly cooperative (with Hill constant greater than 4).(ABSTRACT TRUNCATED AT 250 WORDS)
Shen
L L
LL
Anti-Infective Research Division, Abbott Laboratories, Abbott Park, Illinois 60064.
Baranowski
J
J
Pernet
A G
AG
D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology*; D001667:Binding, Competitive; D004269:DNA, Bacterial / Q000378:metabolism*; D007700:Kinetics; D009643:Norfloxacin / Q000378:metabolism; D011089:Polydeoxyribonucleotides / Q000378:metabolism*; D015363:Quinolones / Q000378:metabolism / Q000494:pharmacology*; D059005:Topoisomerase II Inhibitors
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D011089:Polydeoxyribonucleotides; D015363:Quinolones; D059005:Topoisomerase II Inhibitors; D009643:Norfloxacin