Structural insight into a quinolone-topoisomerase II-DNA complex. Further evidence for a 2:2 quinobenzoxazine-mg2+ self-assembly model formed in the presence of topoisomerase ii.
Quinobenzoxazine A-62176, developed from the antibacterial fluoroquinolones, is active in vitro and in vivo against murine and human tumors. It has been previously claimed that A-62176 is a catalytic inhibitor of mammalian topoisomerase II that does not stabilize the cleaved complex. However, at low drug concentrations and pH 6-7, we have found that A-62176 can enhance the formation of the cleaved complex at certain sites. Using a photocleavage assay, mismatched sequences, and competition experiments between psorospermin and A-62176, we pinpointed the drug binding site on the DNA base pairs between positions +1 and +2 relative to the cleaved phosphodiester bonds. A 2:2 quinobenzoxazine-Mg2+ self-assembly model was previously proposed, in which one drug molecule intercalates into the DNA helix and the second drug molecule is externally bound, held to the first molecule and DNA by two Mg2+ bridges. The results of competition experiments between psorospermin and A-62176, as well as between psorospermin and A-62176 and norfloxacin, are consistent with this model and provide the first evidence that this 2:2 quinobenzoxazine-Mg2+ complex is assembled in the presence of topoisomerase II. These results also have parallel implications for the mode of binding of the quinolone antibiotics to the bacterial gyrase-DNA complex.
Kwok
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Y
Drug Dynamics Institute, College of Pharmacy, University of Texas, Austin, Texas 78712-1074, USA.
Zeng
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Q
Hurley
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LH
D000970:Antineoplastic Agents / Q000378:metabolism*; D020137:Base Pair Mismatch; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D004247:DNA / Q000378:metabolism* / Q000528:radiation effects; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007364:Intercalating Agents / Q000737:chemistry / Q000378:metabolism; D008956:Models, Chemical; D008969:Molecular Sequence Data; D009643:Norfloxacin / Q000378:metabolism; D010078:Oxazines / Q000378:metabolism*; D015363:Quinolones / Q000378:metabolism*; D059005:Topoisomerase II Inhibitors; D014466:Ultraviolet Rays; D014966:Xanthenes / Q000378:metabolism; D044004:Xanthones
D000970:Antineoplastic Agents; D007364:Intercalating Agents; D010078:Oxazines; D015363:Quinolones; D059005:Topoisomerase II Inhibitors; D014966:Xanthenes; D044004:Xanthones; C091925:A 62176; C025543:psorospermin; D004247:DNA; D004250:DNA Topoisomerases, Type II; D009643:Norfloxacin