The adsorption of DNA in the mercury-electrolyte interface

Abstract

AbstractThe adsorption of deoxyribonucleic acid (DNA) in the mercury–electrolyte interface has been investigated. The effect of this adsorption on the differential capacity of the electrical double layer between a polarized mercury surface and an 0.15M NaCl solution containing DNA was measured by means of the alternating current polarography (Breyer polarography). The effective alternating current Ĩ under actual conditions (adsorption processes only, small electrolytic resistance, small alternating current frequency, and alternating current amplitude) is directly proportional to the differential double layer capacity. The combination of this method with the application of a stationary mercury drop electrode allows the coverage of the electrode to be followed, continuously in the range 0.2 sec, to about 60 sec. The diffusion is the rate‐controlled step of the adsorption kinetics. Therefore the lowering of the alternating current Ĩ by the adsorbed DNA is proportional to the surface concentration for partly covered surface and reaches a constant value after the surface becomes fully covered. Adsorption of further layers does not affect the differential capacity. This makes it possible to determine the maximum surface concentration of the DNA. For that it is necessary to determine the diffusion coefficient of DNA. This was done directly by Strassburger and Reinert in our institute. The surface concentrations of the native DNA and the relative surface concentrations of the denatured DNA in dependence on the potential of the polarized mercury surface was estimated. Both surface concentrations show a pronounced dependence on the potential with a minimum of the surface concentration around −0.4 V with respect to the normal calomel electrode. This property may be caused by the structure of the adsorption layer depending on the potential. That means that only several segments at the rigid DNA molecules are adsorbed and the other ones remain in the solution near the surface. The adsorption in the neighborhood of the electrocapillary zero potential at −0.4 V is strongest, and therefore the fraction of the adsorbed segments has a maximum. At these potentials consequently the maximum coverage is already reached at relatively low surface concentrations. Opposite to this is Miller's hypothesis, that native DNA preserves its double helical structure when adsorbed on a negatively charged mercury surface, whereas unfolding occurs on a positively charged mercury surface. Miller's hypothesis is supported by facts that the surface concentration of the denatured DNA should be independent of the potential and should be equal to the surface concentration of the native DNA at a positively charged mercury surface. But an evaluation of Miller's diagrams by no means gives an independence on the potential of the surface concentration of the denatured DNA and no accordance between the surface concentrations of denatured and of native DNA's at the positively charged mercury surface. Moreover Miller compared different DNA samples with different moleculer weights and possibly with different molecular weight distributions. Both the molecular weight and the molecular weight distribution have a pronounced influence on the surface concentration. Therefore this accordance mentioned above is not evident. The critical inspection of Miller's work and the own investigation lead to the conclusion that an unfolding or denaturation of native DNA does not take place in the mercury–electrolyte interface.