Methods to assay inhibitors of DNA gyrase and topoisomerase IV activities.
DNA gyrase and DNA topoisomerase (topo) IV are the bacterial targets of coumarin and quinolone antimicrobial agents. Widespread resistance to clinically important antibiotics such as beta-lactams and macrolides has stimulated the development of novel gyrase and topo IV inhibitors especially against Streptococcus pneumoniae and other Gram-positive pathogens. Here, we describe how gyrase and topo IV activities are measured and how inhibitors of these enzymes may be assayed, focusing as a paradigm on DNA supercoiling by S. pneumoniae gyrase, DNA decatenation by S. pneumoniae topo IV, and DNA cleavage by both enzymes. These approaches provide mechanistic insight on inhibitor action and allow identification of dual gyrase/topo IV targeting agents that can minimize the emergence of bacterial resistance.
Fisher
L Mark
LM
Molecular and Metabolic Signalling Centre, Division of Basic Medical Sciences, St George's, University of London, Cranmer Terrace, London, UK.
Pan
Xiao-Su
XS
D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D002939:Ciprofloxacin / Q000032:analysis / Q000494:pharmacology; D027081:DNA Gyrase / Q000378:metabolism; D027101:DNA Topoisomerase IV / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004269:DNA, Bacterial / Q000032:analysis; D004278:DNA, Superhelical / Q000032:analysis; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000494:pharmacology; D013296:Streptococcus pneumoniae / Q000201:enzymology*; D059005:Topoisomerase II Inhibitors
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004278:DNA, Superhelical; D004791:Enzyme Inhibitors; D059005:Topoisomerase II Inhibitors; D002939:Ciprofloxacin; D027101:DNA Topoisomerase IV; D027081:DNA Gyrase