Amyloid-β Protofibrils: Size, Morphology and Synaptotoxicity of an Engineered Mimic

Abstract

Structural and biochemical studies of the aggregation of the amyloid-β peptide (Aβ) are important to understand the mechanisms of Alzheimer's disease, but research is complicated by aggregate inhomogeneity and instability. We previously engineered a hairpin form of Aβ called Aβcc, which forms stable protofibrils that do not convert into amyloid fibrils. Here we provide a detailed characterization of Aβ42 cc protofibrils. Like wild type Aβ they appear as smooth rod-like particles with a diameter of 3.1 (±0.2) nm and typical lengths in the range 60 to 220 nm when observed by atomic force microscopy. Non-perturbing analytical ultracentrifugation and nanoparticle tracking analyses are consistent with such rod-like protofibrils. Aβ42 cc protofibrils bind the ANS dye indicating that they, like other toxic protein aggregates, expose hydrophobic surface. Assays with the OC/A11 pair of oligomer specific antibodies put Aβ42 cc protofibrils into the same class of species as fibrillar oligomers of wild type Aβ. Aβ42 cc protofibrils may be used to extract binding proteins in biological fluids and apolipoprotein E is readily detected as a binder in human serum. Finally, Aβ42 cc protofibrils act to attenuate spontaneous synaptic activity in mouse hippocampal neurons. The experiments indicate considerable structural and chemical similarities between protofibrils formed by Aβ42 cc and aggregates of wild type Aβ42. We suggest that Aβ42 cc protofibrils may be used in research and applications that require stable preparations of protofibrillar Aβ.

The Alzheimer Disease Protective Mutation A2T Modulates Kinetic and Thermodynamic Properties of Amyloid-β (Aβ) Aggregation

“…6, C and F, and Table 2), considered to be another correlate to the toxic properties of oligomers (37,38). In this assay, the fluorescence of ANS increases upon binding to the lipophilic entities, and the emission maximum undergoes a blue shift.…”

Localization of amyloid beta (Aβ1‐42) protofibrils in membrane lateral compartments: Effect of cholesterol and 7‐Ketocholesterol

“…Fig. S2 shows that assembled species were largely protofibrillar with an average height of 3.18 nm and lengths in the range from 50 to 250 nm [30].…”

Direct High Affinity Interaction between Aβ42 and GSK3α Stimulates Hyperphosphorylation of Tau. A New Molecular Link in Alzheimer’s Disease?

“…9,14 This challenge has been addressed using trapped oligomers, for example, using a disulfide-linked Aβ42 variant that forms relatively large oligomers (protofibrils) that do not convert to fibrils. 15,16 …”