Figure 1.

Oligomerization of Ure2 monitored by confocal single molecule FRET. (A) Schematic figure to indicate the cysteine mutations and fluorescence labeling sites that were used in this study, based on a previously suggested structural model of Ure2 fibrils.⁴⁸ (B) Scheme for smFRET detection of Ure2 oligomers. (C) The concentration of AF555/AF647 labeled Ure2-S68C oligomers throughout the aggregation reaction. (D) Ensemble kinetics of the aggregation of 15 μM (dimeric concentration) unlabeled Ure2-S68C monitored by ThT fluorescence. All the aggregation reactions were carried out at 18 °C in an Innova 4230 incubator with shaking at 150 rpm in 50 mM Tris–HCl (pH 8.4) buffer containing 200 mM NaCl.