Crosslinked α-synuclein for structural assessment was purified as above and additionally immunoprecipitated using the Pierce Direct IP Kit (Thermo Fisher Scientific, Waltham, MA, United States), according to the manufacturer’s instructions. Antibody 2F12 was used as a capture antibody (300 μg per reaction). Volumes of wash and incubation buffers were adapted to the total volume input of the sample. Elution fractions were further concentrated using Amicon concentration columns (Millipore, Burlington, MA, United States) according to the manufacturer’s instructions. The fractions were checked for the purity of α-synuclein multimer using immunoblotting and Coomassie staining. Approximately 10 μM α-synuclein samples were added to a 1 mm path length quartz cuvette for far-UV CD and analyzed using J-1500 CD spectrometer (JASCO) at 25°C. Buffer (50 mM ammonium acetate, pH 7.4) spectra were recorded and subtracted.

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