A monoclonal antibody that inhibits mycobacterial DNA gyrase by a novel mechanism.
DNA gyrase is a DNA topoisomerase indispensable for cellular functions in bacteria. We describe a novel, hitherto unknown, mechanism of specific inhibition of Mycobacterium smegmatis and Mycobacterium tuberculosis DNA gyrase by a monoclonal antibody (mAb). Binding of the mAb did not affect either GyrA-GyrB or gyrase-DNA interactions. More importantly, the ternary complex of gyrase-DNA-mAb retained the ATPase activity of the enzyme and was competent to catalyse DNA cleavage-religation reactions, implying a new mode of action different from other classes of gyrase inhibitors. DNA gyrase purified from fluoroquinolone-resistant strains of M.tuberculosis and M.smegmatis were inhibited by the mAb. The absence of cross-resistance of the drug-resistant enzymes from two different sources to the antibody-mediated inhibition corroborates the new mechanism of inhibition. We suggest that binding of the mAb in the proximity of the primary dimer interface region of GyrA in the heterotetrameric enzyme appears to block the release of the transported segment after strand passage, leading to enzyme inhibition. The specific inhibition of mycobacterial DNA gyrase with the mAb opens up new avenues for designing novel lead molecules for drug discovery and for probing gyrase mechanism.
Manjunatha
Ujjini H
UH
Department of Microbiology and Cell Biology, Indian Institute of Science Bangalore, 560 012, India.
Maxwell
Anthony
A
Nagaraja
Valakunja
V
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism; D027081:DNA Gyrase / Q000378:metabolism; D004278:DNA, Superhelical / Q000378:metabolism; D024881:Drug Resistance, Bacterial; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D009161:Mycobacterium / Q000201:enzymology*; D020102:Mycobacterium smegmatis / Q000187:drug effects / Q000201:enzymology; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000201:enzymology; D021122:Protein Subunits / Q000378:metabolism; D015363:Quinolones / Q000494:pharmacology; D059005:Topoisomerase II Inhibitors
D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D004278:DNA, Superhelical; D004791:Enzyme Inhibitors; D021122:Protein Subunits; D015363:Quinolones; D059005:Topoisomerase II Inhibitors; D004247:DNA; D000251:Adenosine Triphosphatases; D027081:DNA Gyrase