Novel Zn(2+)-chelating peptides selected from a fimbria-displayed random peptide library.
The display of peptide sequences on the surface of bacteria is a technology that offers exciting applications in biotechnology and medical research. Type 1 fimbriae are surface organelles of Escherichia coli which mediate D-mannose-sensitive binding to different host surfaces by virtue of the FimH adhesin. FimH is a component of the fimbrial organelle that can accommodate and display a diverse range of peptide sequences on the E. coli cell surface. In this study we have constructed a random peptide library in FimH. The library, consisting of approximately 40 million individual clones, was screened for peptide sequences that conferred on recombinant cells the ability to bind Zn(2+). By serial selection, sequences that exhibited various degrees of binding affinity and specificity toward Zn(2+) were enriched. None of the isolated sequences showed similarity to known Zn(2+)-binding proteins, indicating that completely novel Zn(2+)-binding peptide sequences had been isolated. By changing the protein scaffold system, we demonstrated that the Zn(2+)-binding seems to be uniquely mediated by the peptide insert and to be independent of the sequence of the carrier protein. These findings might be applied in the design of biomatrices for bioremediation purposes or in the development of sensors for detection of heavy metals.
Kjaergaard
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Microbial Adhesion Group, Section of Molecular Microbiology, BioCentrum-DTU, Technical University of Denmark, Lyngby.
Schembri
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MA
Klemm
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D018829:Adhesins, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018830:Adhesins, Escherichia coli; D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D036781:Fimbriae Proteins; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000378:metabolism*; D010957:Plasmids; D015032:Zinc / Q000378:metabolism*
D018829:Adhesins, Bacterial; D018830:Adhesins, Escherichia coli; D019151:Peptide Library; D010455:Peptides; C058769:fimH protein, E coli; D036781:Fimbriae Proteins; D015032:Zinc