Figure 2. Aggregation kinetics of client proteins in the absence and presence of increasing concentrations of GroEL-AD, as accessed by ThT binding assay.

(a) α-Synuclein; (b) Aβ42 peptide, (c) GroES. For panels (a–c), the red filled circles denote fluorescence values in the absence of GroEL-AD, and the black open symbols denote changes in ThT fluorescence caused by incubation of GroEL-AD alone under identical conditions. The concentration of GroEL-AD added to each experiment was increased according to the following progression of symbols: blue filled squares, green filled diamonds, magenta filled triangles, and orange filled inverted triangles. The specific value of client:GroEL-AD used in each sample (calculated relative to the monomeric molar concentration of client) are as follows in increasing order: (a) 1:0.5, 1:1, 1:2, 1:3; (b) 1:1, 1:5, 1:10, 1:20; (c) 1:0.5, 1:1, 1:2, 1:4. (d) Comparison of the relative effects of GroEL-AD addition on the cumulative fluorescence signal of each client protein. The values are normalized according to the fluorescence values observed for each client protein at the end of the experiment performed in the absence of additional GroEL-AD. The inset to panel (d) is an expansion of the main figure that shows the dependencies at low ratios of GroEL-AD to client.