A streptavidin-metallothionein chimera that allows specific labeling of biological materials with many different heavy metal ions.
We have designed a streptavidin-metallothionein chimeric protein in which the streptavidin moiety provides a means of binding the metallothionein moiety tightly to specific biological targets. A gene fusion of streptavidin with mouse metallothionein I was efficiently expressed in Escherichia coli, and the expressed chimeric protein was purified to homogeneity by a simple procedure. The purified chimera, consisting of four identical subunits, bound one biotin and approximately seven Cd2+ ions per subunit (19.5 kDa). This indicates that both the streptavidin and the metallothionein moieties are fully functional. The high binding affinity of the chimera both for biotin and for heavy metal ions allows the specific labeling or conjugation of any biological material containing unhindered biotin with a variety of different heavy metal ions and their isotopes, thereby opening the way for simultaneous assay systems for a large number of biological targets.
Sano
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Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Glazer
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Cantor
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D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D001483:Base Sequence; D001710:Biotin / Q000737:chemistry; D002104:Cadmium / Q000737:chemistry; D004926:Escherichia coli; D008668:Metallothionein / Q000737:chemistry*; D008670:Metals / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D019809:Streptavidin
D001426:Bacterial Proteins; D008670:Metals; D011993:Recombinant Fusion Proteins; D002104:Cadmium; D001710:Biotin; D019809:Streptavidin; D008668:Metallothionein