title: Designed high affinity Cu2+-binding alpha-helical foldamer.
abstract: The design, synthesis, and characterization of a folded high-affinity metal-binding peptide is described. Based on the previously described folded peptide NTH-18, in which an alpha-helix was constrained through two disulfide bonds to a C-terminal extension of noncanonical secondary structure, a peptide (1) was designed to contain two histidine residues in positions 3 and 7. Air oxidation of 1 led to the formation of peptide 2, which contained two intramolecular disulfide bonds. The presence of the two histidines significantly destabilized the alpha-helical structure of 2 when compared to NTH-18. However, CD spectroscopy revealed that the addition of certain transition metal ions allowed the reformation of a stable alpha-helix. CD, NMR, and EPR spectroscopy as well as MALDI-TOF mass spectrometry indicated that 2 bound to Cu2+ to form a 1:1 complex via the imidazoles of the two histidine side chains. A glycine displacement assay revealed a dissociation constant for this complex of 5 nM at pH 8, which is the lowest reported value for a designed Cu2+-binding peptide. This peptide displayed more than 100-fold selectivity for Cu2+ over Zn2+, Ni2+, and Co2+. The 1.05 A crystal structure of the Cu(II)-complex of 2 revealed a square-pyramidal coordination geometry and confirmed that 2 bound to copper in an alpha-helical conformation via its two histidine side chains. The high affinity metal binding of peptide 2 demonstrates that metals can be used for the selective nucleation of alpha-helices.
authors.affiliation: School of Chemistry, Cardiff University, Main Building, Park Place, Cardiff, CF10 3AT, UK.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Nicoll
authors.forename: David J
authors.initials: DJ
authors.lastname: Miller
authors.forename: Klaus
authors.initials: K
authors.lastname: Fütterer
authors.forename: Raymond
authors.initials: R
authors.lastname: Ravelli
authors.forename: Rudolf K
authors.initials: RK
authors.lastname: Allemann
mesh_terms: D003300:Copper / Q000737:chemistry*; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D017510:Protein Folding; D017433:Protein Structure, Secondary
chemical_list: D010455:Peptides; D003300:Copper