Ligand-Field and Ligand-Binding Analysis of the Active Site of Copper-Bound Aβ Associated with Alzheimer’s Disease

Alzheimer’s disease (AD) patients show abnormally high concentrations of Cu²⁺ in the amyloid β plaques. This invokes that Cu²⁺ might play a crucial role in the onset of AD. The last few decades of research have also shown that Cu²⁺ plays a significant role in the aggregation of Aβ plaques in the brain and the generation of oxidative stress. Because the crystal structures of Cu-Aβ are yet to be obtained, there are various proposed models for the Cu²⁺ coordination environment of Aβ peptides. In this study, we have used the truncated hydrophilic part of the native Aβ peptide to probe the Cu²⁺ coordination site of the peptide, using a combination of spectroscopy and exogenous ligand-binding studies. It is evident from our study that Aβ(1–16) binds 1 equiv of Cu²⁺ and yet shows an equilibrium between two species with a pK_a of ∼8.1. Ligand-field analysis of absorption and circular dichroism spectroscopy data indicates five-coordinate geometry for both components. We investigate the effect of azide and 8-hydroxyquinoline binding to Cu-Aβ and demonstrate the presence of a water-derived ligand and a second exchangeable ligand coordinated to copper at physiological pH, along the equatorial plane of a square-pyramidal active site.