Amyloid β Perturbs Cu(II) Binding to the Prion Protein in a Site-Specific Manner: Insights into Its Potential Neurotoxic Mechanisms

Abstract

Amyloid β (Aβ) is a Cu-binding peptide that plays a key role in the pathology of Alzheimer's disease. A recent report demonstrated that Aβ disrupts the Cu-dependent interaction between cellular prion protein (PrP C ) and N-methyl-D-aspartate receptor (NMDAR), inducing overactivation of NMDAR and neurotoxicity. In this context, it has been proposed that Aβ competes for Cu with PrP C ; however, there is no spectroscopic evidence to support this hypothesis. Prion protein (PrP) can bind up to six Cu(II) ions: from one to four at the octarepeat (OR) region, producing low-and high-occupancy modes, and two at the His96 and His111 sites. Additionally, PrP C is cleaved by αsecretases at Lys110/His111, yielding a new Cu(II)-binding site at the α-cleaved His111. In this study, the competition for Cu(II) between Aβ(1−16) and peptide models for each Cu-binding site of PrP was evaluated using circular dichroism and electron paramagnetic resonance. Our results show that the impact of Aβ(1−16) on Cu(II) coordination to PrP is highly site-specific: Aβ(1−16) cannot effectively compete with the low-occupancy mode at the OR region, whereas it partially removes the metal ion from the high-occupancy modes and forms a ternary OR-Cu(II)-Aβ(1−16) complex. In contrast, Aβ(1−16) removes all Cu(II) ions from the His96 and His111 sites without formation of ternary species. Finally, at the α-cleaved His111 site, Aβ(1−16) yields at least two different ternary complexes depending on the ratio of PrP/ Cu(II)/Aβ. Altogether, our spectroscopic results indicate that only the low-occupancy mode at the OR region resists the effect of Aβ, while Cu(II) coordination to the high-occupancy modes and all other tested sites of PrP is perturbed, by either removal of the metal ion or formation of ternary complexes. These results provide important insights into the intricate effect of Aβ on Cu(II) binding to PrP and the potential neurotoxic mechanisms through which Aβ might affect Cu-dependent functions of PrP C , such as NMDAR modulation.

Water‐Soluble Melanin–Protein–Fe/Cu Conjugates Derived from Norepinephrine as Reliable Models for Neuromelanin of Human Brain Locus Coeruleus

“…These data suggest the presence of essentially square planar copper(II) centers in the Cu‐only conjugates and five‐coordinated copper(II) centers in the Fe‐Cu conjugates [35] . Overall, when inserted into the Peisach–Blumberg diagrams (Figure S7), the parameters suggest that copper(II) displays a mixed nitrogen/oxygen coordination, [36] as it is usually observed for Type 2 sites in copper proteins [37] and in copper(II) complexes with neuronal peptides like amyloid‐β [38–40] and prions, [41, 42] which all display CuN x O y coordination. This indicates that copper(II) is bound exclusively to the protein, βLG or fβLG, and not to the melanin component of the conjugates.…”

Water‐Soluble Melanin–Protein–Fe/Cu Conjugates Derived from Norepinephrine as Reliable Models for Neuromelanin of Human Brain Locus Coeruleus

“…These data suggest the presence of essentially square planar copper(II) centers in the Cu-only conjugates and five-coordinated copper(II) centers in the Fe-Cu conjugates. [35] Overall, when inserted into the Peisach-Blumberg diagrams (Figure S7), the parameters suggest that copper(II) displays a mixed nitrogen/oxygen coordination, [36] as it is usually observed for Type 2 sites in copper proteins [37] and in copper(II) complexes with neuronal peptides like amyloid-β [38][39][40] and prions, [41,42] which all display CuN x O y coordination. This indicates that copper(II) is bound exclusively to the protein, βLG or fβLG, and not to the melanin component of the conjugates.…”

Reversible Stereoisomer‐Specific Cotton Effect of the Ligand Field Transitions at a Copper(II) Binding Site of the Prion Protein

“…The non-octarepeat sites have been recently implicated in the Cu-dependent interaction of PrP with receptors involved in memory and learning. [36] Hence, this knowledge is important to understand the electronic nature of the Cu(II)-His96 complex, and it will probe useful when confronting Cu-PrP complexes to other protein partners.…”