title: Metal binding and folding properties of a minimalist Cys2His2 zinc finger peptide.
abstract: A minimalist Cys2His2 zinc finger peptide, Lys-Tyr-Ala-Cys-Ala-Ala-Cys-Ala-Ala-Ala-Phe-Ala-Ala-Lys-Ala-Ala-Leu-Ala- Ala-His-Ala-Ala-Ala-His-Ala-Lys, has been synthesized. Metal binding studies using Co2+ as a probe indicated that this peptide forms a 1:1 peptide/metal complex with a dissociation constant comparable to that observed for other zinc finger peptides. At high peptide concentrations, a 2:1 peptide/metal complex also forms, with four cysteinates coordinated to Co2+. Additional studies with sequence variants in which the canonical hydrophobic residues were changed to alanine, or in which one of the residues between the cysteines and the histidines was deleted, revealed an even more pronounced formation of the 2:1 complex over the 1:1 complex. In addition, the absorption spectra of the 1:1 peptide/Co2+ complexes of the variant peptides are significantly different from those seen for complexes of the parent peptide or those of more typical zinc finger peptides. NMR studies revealed that the parent peptide folds in the presence of Zn2+ to a structure very similar to that observed for other zinc finger peptides of this class. Taken together, these results suggest that the metal-binding and canonical hydrophobic residues are necessary and sufficient to determine the structure of this class of zinc finger peptides.
authors.affiliation: Department of Chemistry, Johns Hopkins University, Baltimore, MD 21210.
authors.forename: S F
authors.initials: SF
authors.lastname: Michael
authors.forename: V J
authors.initials: VJ
authors.lastname: Kilfoil
authors.forename: M H
authors.initials: MH
authors.lastname: Schmidt
authors.forename: B T
authors.initials: BT
authors.lastname: Amann
authors.forename: J M
authors.initials: JM
authors.lastname: Berg
mesh_terms: D000595:Amino Acid Sequence; D003035:Cobalt / Q000737:chemistry / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D013057:Spectrum Analysis; D013329:Structure-Activity Relationship; D016335:Zinc Fingers
chemical_list: D010455:Peptides; D003035:Cobalt