title: Purification and primary structure of snail metallothionein. Similarity of the N-terminal sequence with histones H4 and H2A.
abstract: A cadmium-binding metallothionein has been purified from metal-exposed Roman snails (Helix pomatia) using gel-permeation, ion-exchange and reverse-phase high-performance liquid chromatography. The S-methylated protein was digested with trypsin and the endoproteinases Asp-N, Glu-C and Arg-C. While most of the resulting peptides could be sequenced by Edman degradation, the intact protein, as well as the N-terminal peptide, proved to be blocked. Analysis by mass spectrometry showed that the N-terminal amino acid was an acetylated serine residue. Snail metallothionein, which is suggested to be involved in the detoxification of cadmium, contains 66 amino acid residues, 18 of which are cysteine residues arranged in seven Cys-Xaa-Cys motifs. The calculated molecular mass of the protein is 6.62 kDa. The primary structure of snail metallothionein reveals a clear relationship with molluscan and vertebrate metallothioneins, but lower similarity with metallothioneins of other invertebrate species. The N-terminal region of the isolated protein proved to be unique among the metallothionein sequences determined so far, showing high degrees of similarity with the N-terminal sequences of histones H2A and H4 which may be important for regulatory functions.
authors.affiliation: Institut für Zoologie (Abteilung Okophysiologie), Universität Innsbruck, Austria.
authors.forename: R
authors.initials: R
authors.lastname: Dallinger
authors.forename: B
authors.initials: B
authors.lastname: Berger
authors.forename: P E
authors.initials: PE
authors.lastname: Hunziker
authors.forename: N
authors.initials: N
authors.lastname: Birchler
authors.forename: C R
authors.initials: CR
authors.lastname: Hauer
authors.forename: J H
authors.initials: JH
authors.lastname: Kägi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002104:Cadmium / Q000378:metabolism; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D006372:Helix, Snails; D006657:Histones / Q000737:chemistry*; D006801:Humans; D008668:Metallothionein / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D017386:Sequence Homology, Amino Acid
chemical_list: D006657:Histones; D002104:Cadmium; D008668:Metallothionein