title: Cloning and specific polymerised-chain-reaction amplification of a third charge-separable human metallothionein isoform.
abstract: Metallothionein (MT) fractions isolated from human adult liver tissue are readily separated by anion-exchange chromatography in two isoforms, MT-1 and MT-2, which differ from each other in the nature of the amino acid residue at position 11. In fetal liver tissue, the presence of a third charge-separable MT isoform has been previously reported. We determined its partial amino acid sequence and the sequence of a cDNA clone encoding this MT form. This confirmed the existence of another human MT isoform, hereafter named MT-0, which is characterized by the presence of a negatively charged amino acid at position 8, and by a Glu23 to Lys substitution in a strictly conserved region of the protein. Taking into account these substitutions, we are able to classify human MT isoforms into three instead of two charge-separable groups, based on the nature of three amino acid residues. The unique presence of Glu8 in MT-0 enabled us to develop an MT-0-specific amplification by the polymerase chain reaction, which revealed the presence of MT-0 mRNA in adult liver RNA samples, in spite of the total absence of this isoform at the protein level. This suggests the involvement of post-transcriptional regulatory mechanisms in the expression of this fetal MT form.
authors.affiliation: Laboratory for Biochemistry, Katholieke Universiteit Leuven, Belgium.
authors.forename: A
authors.initials: A
authors.lastname: Soumillion
authors.forename: J
authors.initials: J
authors.lastname: Van Damme
authors.forename: M
authors.initials: M
authors.lastname: De Ley
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA; D004563:Electrochemistry; D006801:Humans; D008099:Liver / Q000196:embryology / Q000378:metabolism; D008668:Metallothionein / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
chemical_list: D004247:DNA; D008668:Metallothionein