Preferential accumulation of Aβ(1−42) on gel phase domains of lipid bilayers: An AFM and fluorescence study

Peptide–membrane interactions have been implicated in both the toxicity and aggregation of β-amyloid (Aβ) peptides. Recent studies have provided evidence for the involvement of liquid-ordered membrane domains known as lipid rafts in the formation and aggregation of Aβ. As a model, we have examined the interaction of Aβ(1−42) with phase separated DOPC/DPPC lipid bilayers using a combination of atomic force microscopy (AFM) and total internal reflection fluorescence microscopy (TIRF). AFM images show that addition of Aβ to preformed supported bilayers leads to accumulation of small peptide aggregates exclusively on the gel phase DPPC domains. Initial aggregates are observed approximately 90 min after peptide addition and increase in diameter to 45–150 nm within 24 h. TIRF studies with a mixture of Aβ and Aβ–Fl demonstrate that accumulation of the peptide on the gel phase domains occurs as early as 15 min after Aβ addition and is maintained for over 24 h. By contrast, Aβ is randomly distributed throughout both fluid and gel phases when the peptide is reconstituted into DOPC/DPPC vesicles prior to formation of a supported bilayer. The preferential accumulation of Aβ on DPPC domains suggests that rigid domains may act as platforms to concentrate peptide and enhance its aggregation and may be relevant to the postulated involvement of lipid rafts in modulating Aβ activity in vivo.