Polycyclic aromatic hydrocarbons physically intercalate into duplex regions of denatured DNA.
We have investigated the physical binding of pyrene and benzo[a]pyrene derivatives to denatured DNA. These compounds exhibit a red shift in their absorbance spectra of 9 nm when bound to denatured calf thymus DNA, compared to a shift of 10 nm when binding occurs to native DNA. Fluorescence from the hydrocarbons is severely quenched when bound to both native and denatured DNA. Increasing sodium ion concentration decreases binding of neutral polycyclic aromatic hydrocarbons to native DNA and increases binding to denatured DNA. The direct relationship between binding to denatured DNA and salt concentration appears to be a general property of neutral polycyclic aromatic hydrocarbons. Absorption measurements at 260 nm were used to determine the duplex content of denatured DNA. When calculated on the basis of duplex binding sites, equilibrium constants for binding of 7,8,9,10-tetrahydroxy-7,8,9,10-tetrahydro-benzo[a]pyrene to denatured DNA are an order of magnitude larger than for binding to native DNA. The effect of salt on the binding constant was used to calculate the sodium ion release per bound ligand, which was 0.36 for both native and denatured DNA. Increasing salt concentration increases the duplex content of denatured DNA, and it appears that physical binding of polycyclic aromatic hydrocarbons consists of intercalation into these sites.
Division of Toxicology, University of California, San Francisco 94143.
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Wolfe
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Shimer
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Meehan
D004247:DNA; D007364:Intercalating Agents; D007700:Kinetics; D009691:Nucleic Acid Denaturation; D011083:Polycyclic Compounds; D013056:Spectrophotometry, Ultraviolet; D013329:Structure-Activity Relationship
D007364:Intercalating Agents; D011083:Polycyclic Compounds; D004247:DNA