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. 2017 Oct 13;429(20):3018–3030. doi: 10.1016/j.jmb.2017.09.005

Fig. 5.

Fig. 5.

DiY crosslinking stabilizes α-syn fibril seeds. (a) Chemical denaturation in 4 M GdnHCl of WT α-syn fibrils that were (blue) or were not (black) subjected to DiY crosslinking by UV irradiation at 280 nm before, monitored by ThT fluorescence. (b) SEC after 90 min of denaturation, performed on a Superdex 200 column. S: void volume peak containing seeds (> 600 kDa); D: DiY dimer peak; M: α-syn monomer peak. The void volume fractions collected for further characterization are indicated by a blue box. (c) AFM micrograph (height image) of purified DiY-stabilized fibril seeds. (d) Seeding capacity of DiY-stabilized fibril fragments (concentration of 3.5 μM, calculated in monomer units) evaluated by addition to a fibril formation assay of 25 μM non-irradiated α-syn monomer under conditions disfavoring formation of new nuclei or fibril fragmentation (blue). The corresponding SEC fractions of denatured non-crosslinked fibrils (see panel b) were applied as a control (black).