Isolation of recombinant tetrameric N-acetylated α-synuclein

Parkinson's disease (PD) is multifactorial, likely resulting from an intricate relationship of genetic and environmental factors affecting fundamental cellular processes. Histopathological hallmarks of PD include the development of granular inclusions known as Lewy bodies that are enriched with aggregates of the protein α-synuclein (αS). Historically, αS has been considered a natively unfolded protein prone to amyloidogenic behavior. However, recent studies have revealed a physiologically relevant folded αS tetramer that is both alpha-helical and aggregation-resistant. The two forms are thought to reside in a dynamic coexistence within cells, and it has been suggested that a shift from metastable tetramers to the monomeric form could serve as a mechanism for disease initiation. The underlying pathology causing this type of shift remains unknown, but the importance of understanding tetramer stability and disassembly has therapeutic potential that cannot be overemphasized. Isolation of tetrameric αS is complicated by its dynamic nature, so thorough and detailed biochemical and biophysical studies on this αS conformer have been hampered by accessibility issues. We now report a robust and reliable recombinant expression platform that enables purification of native tetrameric αS without any detergents or other structure-modifying additives. [Display omitted] •Native isolation of recombinant noncovalent tetrameric α-synuclein.•Low pressure separation of tetrameric α-synuclein from disordered monomeric form.•Detergent-free recombinant platform for protein complex isolation and enrichment.•Helical α-synuclein tetramer is stable to flash freezing for long-term storage.•Access to tetrameric α-synuclein may lead to novel Parkinson's disease treatments.