Structural and Biophysical Characterization of Stable Alpha-Synuclein Oligomers

The aggregation of alpha-synuclein (alpha-syn) into neurotoxic oligomers and fibrils is an important pathogenic feature of synucleinopatheis, including Parkinson's disease (PD). A further characteristic of PD is the oxidative stress that results in the formation of aldehydes by lipid peroxidation. It has been reported that the brains of deceased patients with PD contain high levels of protein oligomers that are cross-linked to these aldehydes. Increasing evidence also suggests that prefibrillar oligomeric species are more toxic than the mature amyloid fibrils. However, due to the heterogenous and metastable nature, characterization of the alpha-syn oligomeric species has been challenging. Here, we generated and characterized distinct alpha-syn oligomers in vitro in the presence of DA and lipid peroxidation products 4-hydroxy-2-nonenal (HNE) and 4-oxo-2-nonenal (ONE). HNE and ONE oligomer were stable towards the treatment with SDS, urea, and temperature. The secondary structure analysis revealed that only HNE and ONE oligomers contain beta-sheet content. In the seeding assay, both DA and ONE oligomers significantly accelerated the aggregation. Furthermore, all oligomeric preparations were found to seed the aggregation of alpha-syn monomers in vitro and found to be cytotoxic when added to SH-SY5Y cells. Finally, both HNE and ONE alpha-syn oligomers can be used as a calibrator in an alpha-syn oligomers-specific ELISA.

Automated summary

The aggregation of alpha-synuclein into neurotoxic oligomers and fibrils is a key pathological feature in diseases such as Alzheimer's and Parkinson's. This study generated and characterized distinct alpha-synuclein oligomers in vitro in the presence of oxidative stress products. These oligomers were found to be stable and toxic, accelerating alpha-synuclein aggregation and causing cytotoxicity in cells. This research provides valuable insights into the pathogenesis of synucleinopathies and offers potential biomarkers for disease diagnosis.