title: Determination of the drug-DNA binding modes using fluorescence-based assays.
abstract: Therapeutic drugs and environmental pollutants may exhibit high reactivity toward DNA bases and backbone. Understanding the mechanisms of drug-DNA binding is crucial for predicting their potential genotoxicity. We developed a fluorescence analytical method for the determination of the preferential binding mode for drug-DNA interactions. Two nucleic acid dyes were employed in the method: TO-PRO-3 iodide (TP3) and 4',6-diamidino-2-phenylindole (DAPI). TP3 binds DNA by intercalation, whereas DAPI exhibits minor groove binding. Both dyes exhibit significant fluorescence magnification on binding to DNA. We evaluated the DNA binding constant, K(b), for each dye. We also performed fluorescence quenching experiments with 11 molecules of various structures and measured a C(50) value for each compound. We determined preferential binding modes for the aforementioned molecules and found that they bound to DNA consistently, as indicated by other studies. The values of the likelihood of DNA intercalation were correlated with the partition coefficients of the molecules. In addition, we performed nuclear magnetic resonance (NMR) studies of the interactions with calf thymus DNA for the three molecules. The results were consistent with the fluorescence method described above. Thus, we conclude that the fluorescence method we developed provides a reliable determination of the likelihoods of the two different DNA binding modes.
authors.affiliation: Department of Science, John Jay College of Criminal Justice, City University of New York, 445 W 59th St., NY 10019, USA.
authors.forename: Alicia K
authors.initials: AK
authors.lastname: Williams
authors.forename: Sofia Cheliout
authors.initials: SC
authors.lastname: Dasilva
authors.forename: Ankit
authors.initials: A
authors.lastname: Bhatta
authors.forename: Baibhav
authors.initials: B
authors.lastname: Rawal
authors.forename: Melinda
authors.initials: M
authors.lastname: Liu
authors.forename: Ekaterina A
authors.initials: EA
authors.lastname: Korobkova
mesh_terms: D000818:Animals; D001665:Binding Sites; D001681:Biological Assay; D002232:Carbocyanines / Q000737:chemistry; D002417:Cattle; D004247:DNA / Q000737:chemistry*; D004785:Environmental Pollutants / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007202:Indicators and Reagents / Q000737:chemistry; D007211:Indoles / Q000737:chemistry; D007364:Intercalating Agents / Q000737:chemistry*; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D055553:Prescription Drugs / Q000737:chemistry; D013050:Spectrometry, Fluorescence
chemical_list: D002232:Carbocyanines; D004785:Environmental Pollutants; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D007211:Indoles; D007364:Intercalating Agents; D055553:Prescription Drugs; C098830:TO-PRO-3; C007293:DAPI; D004247:DNA; C427203:calf thymus DNA