Preparing Synthetic Aβ in Different Aggregation States

摘要: This chapter outlines protocols that produce homogenous preparations of oligomeric and fibrillar amyloid -β peptide (Aβ). While there are several isoforms this peptide, the 42 amino acid form is focus because its genetic pathological link to Alzheimer’s disease (AD). Past decades AD research highlight dependence Aβ42 function on structural assembly state. Biochemical, cellular in vivo studies usually begin with purified obtained by chemical synthesis or recombinant expression. The initial steps solubilize prepare these dry stocks critical controlling Aβ. To develop assemblies, we initially monomerize erasing any “structural history” could seed aggregation, using a strong solvent. It starting material has allowed us define optimize conditions consistently solutions soluble assemblies. These have been developed characterized atomic force microscopy (AFM) identify structurally discrete species formed under specific solution conditions. used extensively demonstrate variety functional differences between Aβ42. We also present protocol for fluorescently labeling does not affect structure, as measured AFM, function, uptake assay. reagents experimental tools allow defining structure/function connections.