Correlations between subunit distribution, microheterogeneity, and iron content of human liver ferritin.

Subunit heterogeneity of human liver ferritin was investigated by two-dimensional electrophoretic methods. The protein which ordinarily remains assembled in 10 M urea solution was dissociated into subunits in acid-urea or sodium dodecyl sulfate solutions. In agreement with earlier studies, the subunits migrated as two bands in sodium dodecyl sulfate or acid-urea gel electrophoresis systems or in two-dimensional combinations of these systems. Isoelectric focusing methods, however, resolved four major subunit bands and three to five minor bands. Each of these components migrated as either a 22 000 or a 19 000 molecular weight component in sodium dodecyl sulfate gel electrophoresis in the second dimension. The multiple subunit model, which is contrary to currently accepted representations of ferritin structure, is compatible with certain inherent properties of the protein. Thus, ferritin was fractionated on the basis of iron content to show that the relative amounts of individual subunit types were directly dependent upon the iron composition of the protein. Iron-loaded molecules were deficient in the most basic subunit types, and apoferritin was enriched in these components. Aspects of microheterogeneity of assembled ferritin molecules were correlated to subunit heterogeneity, and discrete differences in subunit populations among purified isoferritin components were demonstrated.