Sequential Molecular Interactions Shape Aβ42 Aggregation, Propagation, and Toxicity

Protein aggregation is a context-dependent process in which the molecular environment can influence the properties of the resulting assemblies. In biological systems, these interactions can occur sequentially, as aggregates formed in one cellular or tissue context may encounter different molecular partners and act as seeds in subsequent aggregation events. Here, we used sequential seeding as a controlled experimental model of this temporal and contextual complexity to investigate how prion-like sequences from the gut microbiome modulate amyloid-β aggregation across successive aggregation cycles. Combining kinetic, biophysical, conformational, and toxicity analyses, we show that early interactions with exogenous peptides modify the properties of first-generation Aβ40- and Aβ42-derived seeds, resulting in propagated Aβ42 assemblies with distinct molecular and functional properties. These findings support an Interaction History model in which exogenous sequences bias the emergence of aggregate populations whose properties and subsequent propagation depend on the molecular contexts experienced during earlier aggregation events. Overall, our results present Aβ aggregation as a history-dependent process and suggest that single-step assays may fail to capture aggregate diversity that emerges across successive aggregation cycles.