Characterizing affinity epitopes between prion protein and β-amyloid using an epitope mapping immunoassay

Abstract

Cellular prion protein, a membrane protein, is expressed in all mammals. Prion protein is also found in human blood as an anchorless protein, and this protein form is one of the many potential sources of misfolded prion protein replication during transmission. Many studies have suggested that β-amyloid1–42 oligomer causes neurotoxicity associated with Alzheimer’s disease, which is mediated by the prion protein that acts as a receptor and regulates the hippocampal potentiation. The prevention of the binding of these proteins has been proposed as a possible preventative treatment for Alzheimer’s disease; therefore, a greater understanding of the binding hot-spots between the two molecules is necessary. In this study, the epitope mapping immunoassay was employed to characterize binding epitopes within the prion protein and complementary epitopes in β-amyloid. Residues 23–39 and 93–119 in the prion protein were involved in binding to β-amyloid1–40 and 1–42, and monomers of this protein interacted with prion protein residues 93–113 and 123–166. Furthermore, β-amyloid antibodies against the C-terminus detected bound β-amyloid1–42 at residues 23–40, 104–122 and 159–175. β-Amyloid epitopes necessary for the interaction with prion protein were not determined. In conclusion, charged clusters and hydrophobic regions of the prion protein were involved in binding to β-amyloid1–40 and 1–42. The 3D structure appears to be necessary for β-amyloid to interact with prion protein. In the future, these binding sites may be utilized for 3D structure modeling, as well as for the pharmaceutical intervention of Alzheimer’s disease. Scientists in Korea have identified points of interaction between two key brain proteins that could yield new targets for treating Alzheimer's disease (AD). Cellular prion protein (PrPC) contributes to brain disease if it folds incorrectly, but its normal function in the brain is unknown. Some studies suggest that PrPC mediates the toxic effects of amyloid-β (Aβ), a protein fundamentally associated with AD pathology. To explore this interaction, Mino Kang and Seong Soo A. An of Gachon University, and Su Yeon Kim and Young Ran Ju of the Korea Centers for Disease Control and Prevention, conducted a series of assays to identify how Aβ and PrPC come together. They generated an array of peptides based on short segments of PrPC, and identified multiple regions that specifically bind the Aβ variant associated with AD. Following confirmation of the pathological importance of PrPC-Aβ binding, these interacting sites could offer useful targets for therapeutic interventions.