Tyrosine O-GalNAc Alters the Conformation and Proteolytic Susceptibility of APP Model Glycopeptides

[Image: see text] The amyloid-β precursor protein (APP) undergoes proteolytic cleavage by α-, β-, and γ-secretases, to determine its fate in Alzheimer’s disease (AD) pathogenesis. Recent findings suggest a possible role of O-glycosylation in APP’s proteolytic processing. Therefore, we synthesized native and Swedish-double-mutated APP (glyco)peptides with Tyr(681)-O-GalNAc. We studied conformational changes and proteolytic processing using circular dichroism (CD) spectroscopy and enzyme cleavage assay, respectively. CD analysis was carried out in four solvent systems to evaluate peptide environment and O-glycosylation induced conformational changes. The Swedish mutation and Tyr(681)-O-GalNAc were the key factors driving conformational changes. Furthermore, the level of α- and β-secretase activity was increased by the presence of mutation and this effect was more pronounced for its glycosylated analogues. Our results suggest that O-glycosylation of Tyr(681) can induce a conformational change in APP and affect its proteolytic processing fate toward the amyloidogenic pathway.