Fig. 8: Aggregation kinetics, fibril morphology characterization, and cross-seeding kinetics of single-Ala mutants of Ac1–140. | Nature Communications

Fig. 8: Aggregation kinetics, fibril morphology characterization, and cross-seeding kinetics of single-Ala mutants of Ac1–140.

From: Defining essential charged residues in fibril formation of a lysosomal derived N-terminal α-synuclein truncation

Fig. 8

a Schematic representation of the primary amino acid sequences of Ac1–140 showing sites of mutations. CryoEM fibril core assigned to residues 36 to 99 (gray). Aggregation reactions monitored by ThT fluorescence (10 µM) of 50 µM Ac1–140 (b), AcK80A (c), AcK96A (d) and AcK97A (e) in pH 7.4 buffer (20 mM NaPi, 140 mM NaCl) at 37 °C with continuous linear shaking supplemented with a 2-mm glass bead (n ≥ 4). f Fibril morphologies of single-Ala mutants visualized by TEM. Scale bars are 100 nm. Full-size images are shown in Supplementary Fig. 27. g Aggregation kinetics monitored by ThT fluorescence (10 µM) of soluble Ac1‒140 (50 µM) seeded with 2.5 µM Ac1–140 (black), K80A (blue), K96A (purple) and K97A (green) fibrils in pH 7.4 buffer (20 mM NaPi, 140 mM NaCl) at 37 °C with continuous linear shaking in the absence of 2 mm glass beads. Unseeded 66‒140 (black) is also shown as a control. Solid lines and shaded regions represent mean and SD, respectively (n ≥ 5). Source data are provided as a Source Data file.

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