Abstract: FPLC analysis of the recombinant protein of monkey metallothionein, MT1 and its C33M, C34S and C33M/C34S mutants shows that there were two stable components, f1 and f2. CD study and metal ion analyses indicate that multi components of the mutants relates not only to the binding mode and configuration between cysteins and cadmiun ions, but also to the number of the metal ions. This result also demonstrates that when the Cys33 and Cys34 of α domain of monkey metallothion were mutated into non cystein residues in an attempt of creating a metallothionein with β β domain structure, it led to a protein with multiple structures. It indicates that the α domain plays an important role in the protein stability, and also the distribution of cystein residues affects the formation of metal thiolate clusters and the tertiary structure of the protein significantly.

Key words: Metallothionein, Mutants, FPLC, Circular dichroism spectroscopy, ESI MS, Recombination