Directed molecular screening for RecA ATPase inhibitors.
The roles of bacterial RecA in the evolution and transmission of antibiotic resistance genes make it an attractive target for inhibition by small molecules. We report two complementary fluorescence-based ATPase assays that were used to screen for inhibitors of RecA. We elected to employ the ADP-linked variation of the assay, with a Z' factor of 0.83 in 96-well microplates, to assess whether 18 select compounds could inhibit ATP hydrolysis by RecA. The compounds represented five sets of related inhibitor scaffolds, each of which had the potential to cross-inhibit RecA. Although nucleotide analogs, known inhibitors of GHL ATPases, and known protein kinase inhibitors were not active against RecA, we found that three suramin-like agents substantially inhibited RecA's ATPase activity.
Wigle
Tim J
TJ
School of Pharmacy, Division of Medicinal Chemistry and Natural Products, The University of North Carolina at Chapel Hill, CB #7360, Chapel Hill, NC 27599-7360, USA.
Singleton
Scott F
SF
D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors / Q000378:metabolism; D001665:Binding Sites; D001681:Biological Assay / Q000379:methods; D027081:DNA Gyrase / Q000235:genetics / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000032:analysis / Q000494:pharmacology*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005456:Fluorescent Dyes / Q000737:chemistry*; D006868:Hydrolysis; D009711:Nucleotides / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011938:Rec A Recombinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D012680:Sensitivity and Specificity; D013498:Suramin / Q000032:analysis / Q000494:pharmacology; D013997:Time Factors
D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D009711:Nucleotides; D013498:Suramin; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D011505:Protein-Tyrosine Kinases; D011938:Rec A Recombinases; D000251:Adenosine Triphosphatases; D027081:DNA Gyrase