Studies on mechanism of 8-methoxypsoralen-DNA interaction in the dark

The interaction of 8-methoxypsoralen (8-MOP) with calf thymus DNA was studied in darkness at 25 degrees C and pH 7.4. The enthalpy curve for 8-MOP-DNA interaction was obtained by isothermal titration calorimetry and showed a two-step process for the interaction. According to the spectrophotometric data, it was suggested that some compaction may occur in the DNA structure at higher [8-MOP](t)/[DNA] ratio. Using the fluorescence quenching data, the Scatchard analysis was performed for 8-MOP-DNA interaction at the extended ranges of drug concentration. The results indicated that the first set of binding sites was occupied by 1 mol of drug bound per near eight base pairs of DNA. Also 8-MOP caused the quenching of the fluorescence emission of DNA-ethidium bromide complex. The Scatchard analysis of these data indicated the non-competitive manner for quenching. A non-displacement based quenching mechanism has been suggested for this behavior. The circular dichroism spectra also confirmed the non-intercalative binding of 8-MOP at higher concentrations accompanied by some conformational changes in DNA structure. It has been suggested that at low drug load, 8-MOP binds to DNA as an intercalator, which is an endothermic process, whereas at higher ratios of [8-MOP](t)/[DNA], it binds to the outside of DNA, probably in the minor groove and causes some compaction in DNA, which is the exothermic process.