Transfer of copper from metallothionein to nonmetallothionein proteins in cultured cells

Copper uptake and distribution with time among cytoplasmic proteins were followed in cultured cells under several conditions: (1) CHO cells, which cannot synthesize metallothioneins, were labeled with⁶⁷Cu in the presence of 100 μM ZnCl₂; (2) Cd^r30F9 cells, which contain some constitutive metallothionein (MT), were labeled in the absence of additional ZnCl₂ and; (3) Cd^r30F9 cells were labeled in the presence of ZnCl₂, under which conditions they synthesized additional metallothioneins. The exogenous⁶⁷Cu and ZnCl₂, where present, were then removed, and the distributions of⁶⁷Cu among size fractions of the cellular proteins were observed at intervals for 16 h. In addition, a culture identical to condition (3) above was also treated with 100 μM ZnCl₂ during the redistribution period. The⁶⁷Cu was initially resolved into three peaks by Sephadex G-75 chromatography: high molecular weight, intermediate molecular weight, and MT. The⁶⁷Cu in the MT fraction decreased with at _1/2 of 10–12 h. In contrast to this, generally, in cells with a higher initial⁶⁷Cu bound to metallothionein, there was a progressive increase in the amount of⁶⁷Cu eluting with the high- and intermediate-molecular-weight fractions. Since no other source of⁶⁷Cu was available, these experiments suggest that copper stored in MT can be transferred to other proteins in these cells.