title: Roles of the conserved serines of metallothionein in cadmium binding.
abstract: The sequence of six amino acid residues -Ser-Cys-Cys-Ser-Cys-Cys- is present in all mammalian metallothionein sequences and has been highly conserved during evolution, although the metallothioneins have divergent primary sequences. To determine whether two serines in the sequence play a crucial role in metal-binding of metallothioneins, a mutant metallothionein with these two serines replaced by leucines was obtained using an Escherichia coli expression system. The expressed protein was analyzed for its chemical and spectroscopic properties. It was confirmed that the mutant metallothionein (MT) bound cadmium through a metal-thiolate complex and that there was no strong difference between the mutant and the wild-type MTs in retaining the metal-binding cluster. However, the metal-binding cluster of the mutant metallothionein was more unstable than that of the wild-type metallothionein. The two conservative serines could play a role in the stability of metal-binding ligands.
authors.affiliation: Department of Environmental Medicine and Informatics, Graduate School of Environmental Earth Science, Hokkaido University, Sapporo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Emoto
authors.forename: M
authors.initials: M
authors.lastname: Kurasaki
authors.forename: S
authors.initials: S
authors.lastname: Oikawa
authors.forename: M
authors.initials: M
authors.lastname: Suzuki-Kurasaki
authors.forename: M
authors.initials: M
authors.lastname: Okabe
authors.forename: F
authors.initials: F
authors.lastname: Yamasaki
authors.forename: Y
authors.initials: Y
authors.lastname: Kojima
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002104:Cadmium / Q000378:metabolism*; D017124:Conserved Sequence; D004247:DNA; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D012694:Serine / Q000378:metabolism*
chemical_list: D002104:Cadmium; D012694:Serine; D004247:DNA; D008668:Metallothionein