[Effect of ionic strength of a solution on the protonation of DNA molecule]

The influence of the ionic strength of solution on the DNA molecule protonation was studied by means of circular dichroism (CD), spectrophotometric and potentiometric titration methods over a wide range of the supporting electrolyte concentrations [( NaCl] = 0.0005 divided by 4 M). Consideration of the obtained CD spectra shown that the acidation of the solution induces two cooperative structural transitions in the double stranded DNA molecule in the pre-denaturation pH region. Further decrease in the solution pH results in acidic melting of the DNA molecule. Analysis of the potentiometric data shows that diluted DNA solutions exhibit marked buffer capacity at pH greater than 4.2. A concept of local pH dependent on the electrostatic potential in the vicinity of the polyion was used for interpreting the obtained results. A phase diagram, which describes the polymorphic transformations of the protonated macromolecule, was constructed in terms of pHloc and -log[Na+]. Consideration of this phase diagram allows to hypothesize that: 1) in the neutral diluted DNA solution with a very low supporting electrolyte content the macromolecule exists in a polymorphic state; 2) at [NaCl] greater than or equal to 0.001 M the acid-base equilibrium in the DNA molecule is invariant in respect to the ionic strength of the solution.