Long-range fluorescence quenching of ethidium ion by cationic porphyrins in the presence of DNA
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Abstract
Binding of ethidium to DNA increases the fluorescence efficiency of the molecular ion by about 1 order of magnitude. Quenching of the fluorescence of such an ethidium.DNA complex by tetrakis(4-N-methylpyridyl)porphine (H2T4), and its nickel(II) and zinc(II) derivatives (NiT4 and ZnT4), was investigated in the present study. Results of absorbance and fluorescence measurements obtained for these systems are consistent with a mechanism for quenching that does not involve appreciable displacement of ethidium from the nucleic acid. Sten-Volmer-type plots are nonlinear and can be fit satisfactorily by an equation of the form[GRAPHICS]in which [Q] is the concentration of the porphyrin quencher and sigma is the minimum number of base pairs between ethidium (E+) and the porphyrin required to permit the excited fluorophore to emit a photon. It can be concluded from these studies that, in the presence of DNA, porphyrins are capable of efficiently quenching the fluorescence of an excited ethidium ion at a distance of 25-30 angstrom.
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