Sequestration of cadmium and copper by recombinant rainbow trout and human metallothioneins and by chimeric (mermaid and fishman) proteins with interchanged domains.
A family of synthetic genes was constructed encoding a rainbow trout metallothionein (MT), a human MT, and two chimeric molecules which contained respectively (i) the N-terminal (or head) domain of human MT followed by the C-terminal (or tail) domain of a fish MT (termed mermaid MT) and (ii) the head domain of fish MT fused with the tail domain of human MT (denoted fishman MT). These were expressed in Escherichia coli and the four recombinant proteins were purified to homogeneity therefrom. All four were found to bind 7 g atoms of Cd(II) per mol; at pH 7.0, but not at pH 8.6, four Cd(II) ions were sequestered preferentially in the tail (or alpha) domain. Reciprocally, copper was found to bind preferentially in the head (or beta) domain. The human and fishman MTs displayed a stoichiometry of 12 g atoms of Cu(I) per mol, while rainbow trout and mermaid MTs bound only 10. The significance of these findings is discussed in relation to the different positional organization of cysteine residues close to the N and C termini of mammalian and piscine metallothioneins.
Kille
P
P
Department of Biochemistry, University of Wales College of Cardiff, United Kingdom.
Lees
W E
WE
Darke
B M
BM
Winge
D R
DR
Dameron
C T
CT
Stephens
P E
PE
Kay
J
J
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001059:Apoproteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002104:Cadmium / Q000378:metabolism*; D003300:Copper / Q000378:metabolism*; D003486:Cyanides / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005813:Genes, Synthetic; D006801:Humans; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014337:Trout
D000596:Amino Acids; D001059:Apoproteins; D003486:Cyanides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C031834:apothionein; D002104:Cadmium; D003300:Copper; D008668:Metallothionein