title: Independent self-assembly of cadmium-binding alpha-fragment of metallothionein in Escherichia coli without participation of beta-fragment.
abstract: We examined the independent self-assembly of the alpha- and beta-fragments of human metallothionein (MT) into cadmium-binding conformation in an Escherichia coli expression system, in addition to wild-type MT expression. The expressed alpha-fragment formed independently the structure of a metal-binding cluster without the aid of the beta-fragment. The alpha-fragment and wild-type MT expressed in E.coli were purified and analyzed for their biochemical and spectroscopic properties. The apparent cadmium binding of the alpha-fragment was approximately 12-fold greater than that for the wild-type MT, whereas in other respects the studied biochemical properties were similar. In contrast, we were unable to obtain any independently expressed beta-fragment as the cadmium-binding form in this study. Possible explanations for this phenomenon are discussed.
authors.affiliation: Department of Environmental Medicine and Informatics, Graduate School of Environmental Earth Science, Hokkaido University, Sapporo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kurasaki
authors.forename: T
authors.initials: T
authors.lastname: Emoto
authors.forename: A R
authors.initials: AR
authors.lastname: Arias
authors.forename: M
authors.initials: M
authors.lastname: Okabe
authors.forename: F
authors.initials: F
authors.lastname: Yamasaki
authors.forename: S
authors.initials: S
authors.lastname: Oikawa
authors.forename: Y
authors.initials: Y
authors.lastname: Kojima
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D002104:Cadmium / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D008668:Metallothionein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019216:Metals, Heavy / Q000032:analysis; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D000596:Amino Acids; D019216:Metals, Heavy; D010446:Peptide Fragments; D012333:RNA, Messenger; D011994:Recombinant Proteins; D002104:Cadmium; D008668:Metallothionein