Chromatographic Properties of Metallothioneins on a Gel Permeation Column: Changes Induced by Replacement of Cadmium with Cuprous and Cupric Ions

Abstract

In vitro experiments were conducted to find whether or not a similar elution profile to rat kidney metallothionein with high copper content was obtained on a gel permeation column by replacement of cadmium and/or zinc in rat liver metallothionein with copper. Stepwise replacement of cadmium in rat liver metallothionein with cuprous ion did not cause any shifts of retention times from those of the original proteins on a gel permeation column (SW 3000 column). In contrast to cuprous ion, stepwise replacement of cadmium with cupric ion induced shifts of retention times to larger values than the original ones for the two isometallothioneins on a SW column. Replacement of zinc in zinc-thionein with cupric ion but not with cuprous ion caused a retardation of elution volume on a Sephadex G-75 column. The decreases of cadmium peaks were accompanied by the increases of copper peaks in the case of replacement of cadmium in metallothionein with cuprous ion. Although stepwise decreases of cadmium peaks were observed by the replacement of cadmium in metallothionein with cupric ion, concomitant increases of copper peaks were not observed. Although the relative peak heights of isometallothionein peaks were different from those of rat kidney metallothionein, the third peak with the same retention time as that of rat kidney metallothionein was observed for the replacement of cadmium with cupric but not with cuprous ion.