Solid-phase synthesis and pathological evaluation of pyroglutamate amyloid-beta(3-42) peptide

Pyroglutamate amyloid-beta(3-42) (A beta(pE3-42)) is an N-terminally truncated and pyroglutamate-modified A beta peptide retaining highly hydrophobic, amyloidogenic, and neurotoxic properties. In Alzheimer's disease (AD) patients, A beta(pE3-42) peptides accumulate into oligomers and induce cellular toxicity and synaptic dysfunction. A beta(pE3-42) aggregates further seed the formation of amyloid plaques, which are the pathological hallmarks of AD. Given that A beta(pE3-42) peptides play critical roles in the development of neurodegeneration, a reliable and reproducible synthetic access to these peptides may support pathological and medicinal studies of AD. Here, we synthesized A beta(pE3-42) peptides through the microwave-assisted solid-phase peptide synthesis (SPPS). Utilizing thioflavin T fluorescence assay and dot blotting analysis with anti-amyloid oligomer antibody, the amyloidogenic activity of synthesized A beta(pE3-42) peptides was confirmed. We further observed the cytotoxicity of A beta(pE3-42) aggregates in cell viability test. To examine the cognitive deficits induced by synthetic A beta(pE3-42) peptides, A beta(pE3-42) oligomers were intracerebroventricularly injected into imprinting control region mice and Y-maze and Morris water maze tests were performed. We found that A beta(pE3-42) aggregates altered the expression level of postsynaptic density protein 95 in cortical lysates. Collectively, we produced A beta(pE3-42) peptides in the microwave-assisted SPPS and evaluated the amyloidogenic and pathological function of the synthesized peptides.

Automated summary

A beta(pE3-42) peptides were synthesized using microwave-assisted solid-phase peptide synthesis and their amyloidogenic and pathological functions were evaluated. The synthesized peptides showed amyloidogenic activity and cellular toxicity. In animal models, A beta(pE3-42) aggregates induced cognitive deficits and synaptic dysfunction.