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Biochemistry. 1994 Aug 16;33(32):9699-705.

Formation of mammalian Cu8-metallothionein in vitro: evidence for the existence of two Cu(I)4-thiolate clusters.

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  • 1Biochemisches Institut der Universität Zürich, Switzerland.

Abstract

Copper accumulates in metallothionein (Cu-MT) in copper overload diseases, such as Wilson's disease and Bedlington Terriers disease. The in vitro formation of the Cu12-MT form comprising two Cu(I)6(CysS)9,11 cores is well documented. However, lysosomal Cu-MT isolated from canine liver contains 8 Cu(I) ions in two proposed adamantane-like Cu4-thiolate clusters [Freedman, J. H., Powers, L., & Peisach, J. (1986) Biochemistry 25, 2342]. The present studies have been carried out in an effort to learn more about the Cu(I)-thiolate cluster species formed upon the sequential incorporation of Cu(I) ions into metal-free MT from rabbit liver. On the basis of changes in the electronic absorption, circular dichroism (CD), magnetic circular dichroism (MCD), and luminescence spectra, besides the formation of a molecular species with 12 Cu(I) equivalents, evidence for the existence of a distinct MT complex with 8 Cu(I) equivalents (Cu8-MT) was obtained. Analysis of the metal-dependent absorption envelope of Cu(I)-MT between 240 and 360 nm permitted the discrimination between predominantly CysS-Cu(I) charge-transfer (LMCT) (240-260 nm) and cluster-localized Cu(I) (d-s) transitions (260-360 nm). Accordingly, the decrease in the ratio of intensities of LMCT to d-s bands from 2.6 to 2.4 on going from 8 to 12 Cu(I) equivalents was attributed to the formation of Cu-MT species with different cysteine ligand to metal stoichiometries. The results suggest that while in Cu12-MT all 20 thiolate ligands participate in metal binding, in the Cu8-MT species between 12 and 14 cysteines take part in Cu(I) coordination.(ABSTRACT TRUNCATED AT 250 WORDS)

PMID:
8068648
[PubMed - indexed for MEDLINE]
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