Activities of fluoroquinolones against Streptococcus pneumoniae type II topoisomerases purified as recombinant proteins.
Streptococcus pneumoniae topoisomerase IV and DNA gyrase have been purified from a fluoroquinolone-susceptible Streptococcus pneumoniae strain, from first-step mutants showing low-level resistance to ciprofloxacin, sparfloxacin, levofloxacin, and ofloxacin, and from two clinical isolates showing intermediate- and high-level fluoroquinolone resistance by a gene cloning method that produces recombinant proteins from Escherichia coli. The concentrations of ciprofloxacin, sparfloxacin, levofloxacin, or ofloxacin required to inhibit wild-type topoisomerase IV were 8 to 16 times lower than those required to inhibit wild-type DNA gyrase. Furthermore, low-level resistance to these fluoroquinolones was entirely due to the reduced inhibitory activity of fluoroquinolones against topoisomerase IV. For all the laboratory strains, the 50% inhibitory concentration for topoisomerase IV directly correlated with the MIC. We therefore propose that with S. pneumoniae, ciprofloxacin, sparfloxacin, levofloxacin, and ofloxacin target topoisomerase IV in preference to DNA gyrase. Sitafloxacin, on the other hand, was found to be equipotent against either enzyme. This characteristic is unique for a fluoroquinolone. A reduction in the sensitivities of both topoisomerase IV and DNA gyrase are required, however, to achieve intermediate- or high-level fluoroquinolone resistance in S. pneumoniae.
Morrissey
I
I
GR Micro Ltd., London NW1 3ER, United Kingdom. i.morrissey@grmicro.co.uk
George
J
J
D000890:Anti-Infective Agents / Q000494:pharmacology*; D027101:DNA Topoisomerase IV; D004250:DNA Topoisomerases, Type II / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000302:isolation & purification / Q000378:metabolism; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000378:metabolism; D024841:Fluoroquinolones; D008826:Microbial Sensitivity Tests; D009154:Mutation; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification; D013296:Streptococcus pneumoniae / Q000187:drug effects / Q000201:enzymology*; D059005:Topoisomerase II Inhibitors
D000890:Anti-Infective Agents; D004269:DNA, Bacterial; D024841:Fluoroquinolones; D011994:Recombinant Proteins; D059005:Topoisomerase II Inhibitors; D027101:DNA Topoisomerase IV; D004250:DNA Topoisomerases, Type II