Interaction of ethidium bromide and caffeine with DNA in aqueous solution |
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Authors: | S F Baranovsky P A Bolotin M P Evstigneev and D N Chernyshev |
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Institution: | (1) Sevastopol National Technical University, Studgorodok, Sevastopol, 99053, Ukraine |
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Abstract: | Two component (ethidium bromide–caffeine, ethidium bromide–DNA) and three component (ethidium bromide–caffeine–DNA) systems
in aqueous saline (0.01 M NaCl) phosphate buffer solutions (pH 6.86, T = 298 K) are studied spectrophotometrically. The equilibrium constants for dimerization of caffeine, K
D
= 1.22 ± 2 M−1, and for heteroassociation of ethidium bromide with caffeine, K = 71 ± 8 M−1, in ethidium bromide–caffeine systems are determined. When the concentration of caffeine is increased, the dynamic equilibrium
of the solution shifts toward formation of heterocomplexes which are, presumably, stabilized by dispersive and hydrophobic
interactions of chromophores. The equilibrium parameters for ethidium bromide complex formation with DNA are calculated: the
coupling constant for the dye with the biopolymer, K
1 = (232 ± 16)⋅103 M−1, and the number of base pairs of the biopolymer participating in bonding with the ligand, n
1 = 3.6 ± 0.2, are calculated. Given these values, it is suggested that under these experimental conditions there are two types
of bonding between ethidium bromide and the nucleic acid — intercalation and “external” bonds. A McGhee–von Hippel model for
a three component system and the numerical values of the parameters for molecular complex formation in two component systems
are used to calculate the bonding constant for caffeine with DNA, K
2 = 127 ± 30 M−1, and the number of base pairs of DNA which bond with caffeine, n
2 = 1.7 ± 0.2. The concentrations of ethidium bromide and caffeine in the composition of two and three component complexes
are calculated as functions of the nucleic acid content in the solution. An analysis of the concentration dependences shows
that heteroassociation of ligands has a significant effect on the reduction in the concentration of ethidium bromide–DNA complexes
in a three component system for low DNA concentrations, while at high DNA concentrations the bonding of caffeine with the
biopolymer has this effect.
Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 76, No. 1, pp. 143–151, January–February 2009. |
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Keywords: | spectrophotometry optical density ethidium bromide caffeine ligand complex formation heteroassociation dimerization constant DNA complex formation constant McGhee-von Hippel model intercalation bonding “ external” bonding |
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