Metal co-ordination in rat liver metallothionein-2 prepared with or without reconstitution of the metal clusters, and comparison with rabbit liver metallothionein-2.
Possible origins of the different metal co-ordination topologies in the recently determined structures of rat metallothionein-2 (MT2) in single crystals and rabbit MT2 in solution were investigated. A complete structure determination for rat MT2 in solution by nuclear magnetic resonance (n.m.r.) showed that the differences in the spatial structures cannot be attributed to the different primary structures of the two species. Comparison of [113Cd7]MT2 obtained by reconstitution of the apoprotein in vitro with preparations using a different procedure showed, moreover, that the metal co-ordination observed in solution by n.m.r. is not an artefact of the protein reconstitution. Solutions of high-pressure liquid chromatographically homogeneous biosynthetic preparations of [113Cd, Zn]MT2 were obtained from rat liver following injection of 113Cd into rats in vivo, without further metal exchange after protein isolation. They contain a mixture of several forms of MT2 with different relative metal compositions, giving rise to an increased number of 113Cd resonances. For the components of the four-metal cluster, the major one of these different forms exhibits patterns in the two-dimensional [1H, 113Cd]-correlated spectra that are indistinguishable from those of [113Cd7]MT2, thereby implying identity of cluster coordination and topology. These results are discussed with regard to continued investigations into the differences between the solution structure and crystal structure of MT2.
Vasák
M
M
Biochemisches Institut, Universität Zürich, Switzerland.
Wörgötter
E
E
Wagner
G
G
Kägi
J H
JH
Wüthrich
K
K
D000595:Amino Acid Sequence; D000818:Animals; D002104:Cadmium; D008099:Liver / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008668:Metallothionein; D008969:Molecular Sequence Data; D011817:Rabbits; D051381:Rats; D015032:Zinc
D002104:Cadmium; D008668:Metallothionein; D015032:Zinc