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Chembiochem. 2014 Apr 14;15(6):822-825. doi: 10.1002/cbic.201300727. Epub 2014 Mar 3.

Evolution of iron(II)-finger peptides by using a bipyridyl amino acid.

Author information

1
Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Rd, La Jolla, CA 92037.
2
Department of Chemistry, Stanford University, 333 Campus Drive, Stanford CA 94305.
#
Contributed equally

Abstract

We report the engineering of zinc-finger-like motifs containing the unnatural amino acid (2,2'-bipyridin-5-yl)alanine (Bpy-Ala). A phage-display library was constructed in which five residues in the N-terminal finger of zif268 were randomized to include both canonical amino acids and Bpy-Ala. Panning of this library against a nine-base-pair DNA binding site identified several Bpy-Ala-containing functional Zif268 mutants. These mutants bind the Zif268 recognition site with affinities comparable to that of the wild-type protein. Further characterization indicated that the mutant fingers bind low-spin Fe(II) rather than Zn(II) . This work demonstrates that an expanded genetic code can lead to new metal ion binding motifs that can serve as structural, catalytic, or regulatory elements in proteins.

KEYWORDS:

amino acids; bipyridyl complexes; iron; phage display; zinc finger engineering

PMID:
24591102
PMCID:
PMC4010245
DOI:
10.1002/cbic.201300727
[Indexed for MEDLINE]
Free PMC Article

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