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. 2011 Jul 11;194(1):49–60. doi: 10.1083/jcb.201010117

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© 2011 Krumova et al.

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Figure 2. — In vitro fibril formation of sumoylated α-synuclein. (A) Coomassie staining of α-synuclein sumoylated to 100% (lane 1) and α-synuclein with an equimolar amount of free SUMO1 (lane 2). (B) Fibrillization kinetics of α-synuclein sumoylated to 100% (70 µM; closed circles) and control condition (70 µM of nonmodified α-synuclein + 70 µM of free SUMO1; open circles). (C) Nonamyloidogenic amorphous oligomers formed by α-synuclein sumoylated to 100%. (D) Mature fibrils formed by control nonmodified α-synuclein in the presence of 70 µM of free SUMO1. (E) Coomassie staining of α-synuclein sumoylated to 50% (lane 1) and α-synuclein with an equimolar amount of free SUMO1 (lane 2). (F) Fibrillization kinetics of α-synuclein sumoylated to 50% (35 µM sumoylated α-synuclein + 35 µM of nonmodified α-synuclein; closed circles) and control condition (70 µM of nonmodified α-synuclein + 35 µM of free SUMO1; open circles). (G) Nonamyloidogenic amorphous material formed by α-synuclein sumoylated to 50%. (H) Mature fibrils formed by control nonmodified α-synuclein in the presence of 35 µM of free SUMO1. (I) Coomassie staining of α-synuclein sumoylated to 10% (lane 1) and α-synuclein with an equimolar amount of free SUMO1 (lane 2). (J) Fibrillization kinetics of α-synuclein sumoylated to 10% (7 µM sumoylated α-synuclein + 63 µM of nonmodified α-synuclein; closed circles) and control condition (70 µM of nonmodified α-synuclein + 7 µM of free SUMO1; open circles). (K) Mature fibrils formed by α-synuclein sumoylated to 10%. (L) Mature fibrils formed by control nonmodified α-synuclein in the presence of 7 µM of free SUMO1. (C, D, G, H, K, and L) TEM of aggregation samples after 146 h of incubation in 50 mM Hepes and 100 mM NaCl, pH 7.4, at 37°C with constant stirring. ThT, Thioflavin T. Bars, 200 nm.