Aggregation studies of de-metallated and reduced SOD1-PB site specific mutants on treatment with PB.

ThT fluorescence was monitored during the co-incubation of (a) SOD1 A123F (50 μM) with PB at 1:30 molar ratio (b) in the presence of 5mM EDTA, (c) SOD1 K9F (50 μM) with PB at 1:30 molar ratio (d) in the presence of 5mM EDTA (e) SOD1 G10P (50 μM) with PB at 1:30 molar ratio (f) in the presence of 5mM EDTA (g) SOD1 A123F (50 μM) with PB at 1:30 molar ratio (h) in the presence of 5mM EDTA, at 37 °C with continuous shaking, under reduced conditions. The values were normalized to the maximal ThT intensity and fitted to a Boltzmann sigmoidal equation. The control and PB-treated species were compared to analyse PB’s effects on the aggregation of SOD1 and SOD1. All the experiments were performed for n = 3 biological replicates. (DOCX)