Structure of the copper cluster in canine hepatic metallothionein using X-ray absorption spectroscopy.

The metal binding site in the lysosomal copper metallothionein from canine liver (LyCuLP) was examined with X-ray edge and extended X-ray absorption fine structure (EXAFS) spectroscopies. The k-absorption edge spectrum of LyCuLP was consistent with the coordination of univalent copper. The Fourier transform of the EXAFS data showed four resolved shells of backscattering atoms. Comparisons between the phase and amplitude functions derived from the isolated shells to those of Cu-Cu, Cu-S, and Cu-N model compounds showed that each copper was coordinated by four sulfur atoms at a distance of 2.27 +/- 0.02 A. Analysis of the outer shell data indicated backscattering copper atoms at 2.74 +/- 0.05, 3.32 +/- 0.05, and 3.88 +/- 0.05 A. Interatomic distances determined from the EXAFS data were compared to the distances observed by X-ray crystallographic analysis of adamantane-like clusters containing four and five copper atoms and a cubic cluster containing four copper atoms, structurally similar to the 4Fe-4S clusters in some ferredoxins. The results of these comparisons suggest that the copper complexed in LyCuLP is arranged in an adamantane-like cluster. The structure derived for this protein may be conserved in other copper metallothioneins.