AN ELECTROPHORETIC STUDY OF MIXTURES OF OVALBUMIN AND YEAST NUCLEIC ACID

Abstract

Electrophoretic patterns of mixtures of ovalbumin and yeast nucleic acid indicate that the constituents migrate independently of each other in buffer solutions of 0.1 ionic strength and at pH values somewhat higher than the isoelectric point of the protein. In the isoelectric region, however, the patterns from the two sides of the channel exhibit asymmetries that can be explained by assuming the existence in the mixture of appreciable concentrations of a reversibly dissociable complex between the components. Formation of this complex is favored by increasing concentrations of the components and decreasing ionic strength. At pH values below the isoelectric point partial precipitation of the complex occurs. The patterns obtained from each side of the channel in the electrophoresis of a mixture of two components, which form a dissociable complex, indicate only two boundaries, aside from the δ and ϵ effects. One of these is a normal boundary whose displacement is proportional to the mobility of a component that has separated from the mixture. In the other boundary, however, dissociation of the complex occurs and consequently the displacement of this boundary corresponds to the mobility of neither component nor to that of the complex. Moreover, the areas under the refractive index gradient curves are not proportional to the stoichiometric concentrations of the components. However, equations are developed with the aid of which an electrophoretic analysis of the mixture is possible. This analysis requires the use of data from the patterns of both channels.