Coordination of three and four Cu(I) to the α− and β‐domain of vertebrate Zn‐metallothionein‐1, respectively, induces significant structural changes

Vertebrate metallothioneins are found to contain Zn(II) and variable amounts of Cu(I), in vivo, and are believed to be important for d10‐metal control. To date, structural information is available for the Zn(II) and Cd(II) forms, but not for the Cu(I) or mixed metal forms. Cu(I) binding to metallothionein‐1 has been investigated by circular dichroism, luminescence and 1H NMR using two synthetic fragments representing the α‐ and the β‐domain. The 1H NMR data and thus the structures of Zn4α metallothionein (MT)‐1 and Zn3βMT‐1 were essentially the same as those already published for the corresponding domains of native Cd7MT‐1. Cu(I) titration of the Zn(II)‐reconstituted domains provided clear evidence of stable polypeptide folds of the three Cu(I)‐containing α‐ and the four Cu(I)‐containing β‐domains. The solution structures of these two species are grossly different from the structures of the starting Zn(II) complexes. Further addition of Cu(I) to the two single domains led to the loss of defined domain structures. Upon mixing of the separately prepared aqueous three and four Cu(I) loaded α‐ and β‐domains, no interaction was seen between the two species. There was neither any indication for a net transfer of Cu(I) between the two domains nor for the formation of one large single Cu(I) cluster involving both domains.