Stabilization of α-synuclein oligomers using formaldehyde

Autoři: Harm Ruesink aff001;  Lasse Reimer aff001;  Emil Gregersen aff001;  Arne Moeller aff001;  Cristine Betzer aff001;  Poul Henning Jensen aff001
Působiště autorů: Danish Research Institute of Translational Neuroscience—DANDRITE, Aarhus University, Aarhus, Denmark aff001;  Department of Biomedicine, Aarhus University, Aarhus, Denmark aff002;  Department of Structural Biology, The Max Planck Institute of Biophysics, Frankfurt, Germany aff003
Vyšlo v časopise: PLoS ONE 14(10)
Kategorie: Research Article
doi: 10.1371/journal.pone.0216764


The group of neurodegenerative diseases, Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA) all exhibit inclusions containing amyloid-type α-synuclein (α-syn) aggregates within degenerating brain cells. α-syn also exists as soluble oligomeric species that are hypothesized to represent intermediates between its native and aggregated states. These oligomers are present in brain extracts from patients suffering from synucleinopathies and hold great potential as biomarkers. Although easily prepared in vitro, oligomers are metastable and dissociate over time, thereby complicating α-syn oligomer research. Using the small amine-reactive cross-linker, formaldehyde (FA), we successfully stabilized α-syn oligomers without affecting their size, overall structure or antigenicity towards aggregate-conformation specific α-syn antibodies FILA and MJFR-14-6-4-2. Further, cross-linked α-syn oligomers show resistance towards denaturant like urea and SDS treatment and remain fully functional as internal standard in an aggregation-specific enzyme-linked immunosorbent assay (ELISA) despite prior incubation with urea. We propose that FA cross-linked α-syn oligomers could serve as important calibrators to facilitate comparative and standardized α-syn biomarker studies going forward.

Klíčová slova:

Amyloid proteins – Antigens – Cross-linking – Enzyme-linked immunoassays – Monomers – Oligomers – Parkinson disease – Formaldehyde


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2019 Číslo 10