Structural Insight into Redox Dynamics of Copper Bound N-Truncated Amyloid-β Peptides from in Situ X-ray Absorption...

X-ray absorption spectroscopy of Cu^II amyloid-β peptide (Aβ) under in situ electrochemical control (XAS-EC) has allowed elucidation of the redox properties of Cu^II bound to truncated peptide forms. The Cu binding environment is significantly different for the Aβ_1-16 and the N-truncated Aβ_4-9, Aβ_4-12, and Aβ_4-16 (Aβ_4-9/12/16) peptides, where the N-truncated sequence (F₄R₅H₆) provides the high-affinity amino-terminal copper nickel (ATCUN) binding motif. Low temperature (ca. 10 K) XAS measurements show the adoption of identical Cu^II ATCUN-type binding sites (Cu^II_ATCUN) by the first three amino acids (FRH) and a longer-range interaction modeled as an oxygen donor ligand, most likely water, to give a tetragonal pyramid geometry in the Aβ_4-9/12/16 peptides not previously reported. Both XAS-EC and EPR measurements show that Cu^II:Aβ_4-16 can be reduced at mildly reducing potentials, similar to that of Cu^II:Aβ_1-16. Reduction of peptides lacking the H₁₃H₁₄ residues, Cu^II:Aβ_4-9/12, require far more forcing conditions, with metallic copper the only metal-based reduction product. The observations suggest that reduction of Cu^II_ATCUN species at mild potentials is possible, although the rate of reduction is significantly enhanced by involvement of H₁₃H₁₄. XAS-EC analysis reveals that, following reduction, the peptide acts as a terdentate ligand to Cu^I (H₁₃, H₁₄ together with the linking amide oxygen atom). Modeling of the EXAFS is most consistent with coordination of an additional water oxygen atom to give a quasi-tetrahedral geometry. XAS-EC analysis of oxidized Cu^II:Aβ_4-12/16 gives structural parameters consistent with crystallographic data for a five-coordinate Cu^III complex and the Cu^II_ATCUN complex. The structural results suggest that Cu^II and the oxidation product are both accommodated in an ATCUN-like binding site.