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PMID: 2831810 Published · ppublish English Journal Article

Purification of Citrobacter freundii DNA gyrase and inhibition by quinolones.

Antimicrobial agents and chemotherapy ·Vol. 32 ·No. 1 ·1988-01-00 ·Pages 104-9

Aoyama H, Sato K, Fujii T, Fujimaki K, Inoue M, Mitsuhashi S

Abstract

DNA gyrase is a bacterial enzyme which catalyzes the ATP-dependent negative supercoiling of DNA. It is the accepted target of quinolones. The enzyme from Citrobacter freundii IID976 was purified by affinity chromatography on novobiocin-Sepharose and heparin-Sepharose. It had two subunits, designated A and B, which closely resembled those of the enzyme from Escherichia coli and Micrococcus luteus in enzymatic requirements. The inhibitory effects of the quinolones on the supercoiling activities of the enzyme correlated with their antibacterial activities. New quinolones were better inhibitors of DNA gyrase than nalidixic acid and pipemidic acid. We also purified DNA gyrase from a spontaneous nalidixic acid-resistant mutant (M2-5). The gyrases from IID976 and M2-5 were defined as mixtures of subunits As+Bs (s, susceptible) and Ar+Br (r, resistant), respectively. The supercoiling activities of reconstituted Ar+Br and Ar+Bs were more resistant to quinolones than As+Bs and As+Br. These findings indicate that one mechanism of C. freundii resistance against quinolones is resistance modification of the A subunit protein.

MeSH Terms
Anti-Bacterial Agents/pharmacology Citrobacter/drug effects,enzymology,metabolism DNA Topoisomerases, Type II/isolation & purification Drug Resistance, Microbial Nalidixic Acid/pharmacology Norfloxacin/metabolism,pharmacology Quinolines/pharmacology Topoisomerase II Inhibitors
Chemicals
Anti-Bacterial Agents Quinolines Topoisomerase II Inhibitors Nalidixic Acid DNA Topoisomerases, Type II Norfloxacin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Aoyama H
Episome Institute, Seta-gun, Japan.
Sato K
Fujii T
Fujimaki K
Inoue M
Mitsuhashi S
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Article Info
Journal
Antimicrobial agents and chemotherapy
Abbr.
Antimicrob Agents Chemother
ISSN
0066-4804
DOI
10.1128/AAC.32.1.104
Published
1988-01-00
Pages
104-9
Language
English
Region
United States
NLM ID
0315061
PMCID
PMC172107
Subset
IM
External Links
PubMed source DOI: 10.1128/AAC.32.1.104 PMC Full Text View journal details
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