Differential pulse voltammetric studies of ethidium bromide binding to DNA.

The interaction of ethidium bromide (EtBr) with calf thymus DNA is investigated electrochemically with the use of differential pulse voltammetry (DPV) at two different ionic strengths of a solution (0.154 M and 0.02 M [Na+], pH 7.0). It is revealed that EtBr binds with DNA in more than one way. The appropriate values of constants (K) and number site sizes (n) of EtBr binding to DNA are determined. The values of binding constants are equal to 1.9 x 10(6) and 5.6 x 10(5) M(-1), and number site sizes to 9 and 3.6 for strong interactions at ionic strengths of solutions 0.02 and 0.154 M Na+ at 28 degrees C, respectively. For a weaker interaction, these parameters are equal to 7 x 10(4) and 8 x 10(4) M(-1) and 1.5 and 1 at the mentioned ionic strengths of solutions, respectively. Thus, EtBr interacts with DNA in more than one way--intercalative and electrostatic at low ionic strength, and semi-intercalative and electrostatic at a higher strength of the solution. These results are in good accordance with the ones obtained by spectroscopic (absorption and fluorimetric) methods.

Kinetics of ligand binding to nucleic acids.

Ligand binding to nucleic acids (NA) is considered as a stationary Markov process. It is shown that the probabilistic description of ligand-NA binding allows one to describe not only the kinetics of the change of number of bound ligands at arbitrary fillings but also to calculate stationary values of the number of bound ligands and its dispersion. The general analysis of absorption isotherms and kinetics of ligand binding to NA make it possible to determine of rate constants of ligand-NA complex formation and dissociation.

The binding of ethidium bromide with DNA: interaction with single- and double-stranded structures.

The pH-induced helix-coil transition of DNA and its complexes with EtBr is carried out at acidic pH in a wide interval of change of concentration ratio of EtBr/DNA. The binding isotherms of EtBr on double and single-stranded DNA at pH = 7.0 and pH = 3.0 (t = 25(o)C) are obtained by absorption and fluorimetric methods. Binding constants (K) and number of bases (n), corresponding to one binding site were determined. Non fluorescent "strong" complex with ds-DNA at pH = 7.0 and t = 25(o)C as well as "strong" and "weak" complexes with ss-DNA at pH = 3.0 and t = 25(o)C are revealed.

DNA structure properties and phospholipid composition of Salmonella derby.

The properties of DNA structure and the phospholipid content of Salmonella derby cells were studied with respect to their plasmid content and radiosensitivity. The role of R-plasmid in determining the qualitative and quantitative compositions of S. derby phospholipids was revealed. The radiosensitivity of plasmid-carrying S. derby mutants was shown to be most likely determined by the structure of DNA, its GC content, and the level of methylation. We suggest that the phospholipid molecules and their interaction with DNA play a key role in formation of the radio-resistance of plasmid-free S. derby cells.