The Role of Cholesterol in Amyloidogenic Substrate Binding to the γ-Secretase Complex

Alzheimer’s disease is the most common progressive neurodegenerative disorder and is characterized by the presence of amyloid β (Aβ) plaques in the brain. The γ-secretase complex, which produces Aβ, is an intramembrane-cleaving protease consisting of four membrane proteins. In this paper we investigated the amyloidogenic fragments of amyloid precursor protein (substrates Aβ₄₃ and Aβ₄₅, leading to less amyloidogenic Aβ₄₀ and more amyloidogenic Aβ₄₂, respectively) docked to the binding site of presenilin, the catalytic subunit of γ-secretase. In total, we performed 9 μs of all-atom molecular dynamics simulations of the whole γ-secretase complex with both substrates in low (10%) and high (50%) concentrations of cholesterol in the membrane. We found that, at the high cholesterol level, the Aβ₄₅ helix was statistically more flexible in the binding site of presenilin than Aβ₄₃. An increase in the cholesterol concentration was also correlated with a higher flexibility of the Aβ₄₅ helix, which suggests incompatibility between Aβ₄₅ and the binding site of presenilin potentiated by a high cholesterol level. However, at the C-terminal part of Aβ₄₅, the active site of presenilin was more compact in the case of a high cholesterol level, which could promote processing of this substrate. We also performed detailed mapping of the cholesterol binding sites at low and high cholesterol concentrations, which were independent of the typical cholesterol binding motifs.