Data associated with "Defining essential charged residues in fibril formation of a lysosomal derived N-terminal alpha-synuclein truncation

Source data:

• Figure 1d-g: Activity assays for CtsB, CtsL, CtsD and AEP
• Figure 2a: ThT aggregation kinetic data for Ac1-140 and 66-140
• Figure 2c and S6: Raman data on Ac1-140 and 66-140 fibrils
• Figure 3a: 2D ¹³C-¹³C ssNMR data for 66-140 fibrils
• Figure 3b: Secondary chemical shift data for 66-140
• Figure 4b and S14: ThT aggregation kinetic data for 66-140 and single ala-mutants
• Figure 5a and S16: Raman data 66-140 and single ala-mutant
• Figure 5b: Raman difference data subtracting 66-140 from single ala-mutants
• Figure 5c: ThT aggregation kinetic data of soluble 66-140 seeded with single ala-mutant fibrils
• Figure 6a: ThT aggregation kinetic data of single ala-mutants seeded with 66-140 fibrils
• Figure 6c and S22: Raman data of single-Ala mutants seeded with 66–140 fibrils
• Figure 7a: ThT aggregation kinetic data of charge switch mutants seeded with 66-140 fibrils
• Figure 7c and S24: Raman data of E83K/K97E seeded with 66–140 fibrils
• Figure 8b: ThT aggregation kinetic data for Ac1-140
• Figure 8c: ThT aggregation kinetic data AcK80A
• Figure 8d: ThT aggregation kinetic data AcK96A
• Figure 8e: ThT aggregation kinetic data AcK97A
• Figure 8g: ThT aggregation kinetic data of soluble Ac1-140 seeded with single-Ala full-length mutants
• Figure S3: Data for a salt-dependent aggregation of 66–140
• Figure S4a: ThT aggregation kinetic data for self-seeding 66‒140
• Figure S7a: 2D carbon-carbon cross-polarization data
• Figure S7b: Data for 1D slices along the ¹³C dimension at indicated positions in 2D carbon-carbon spectrum
• Figure S8a: 2D NCACX data on 66‒140 fibrils
• Figure S8b: Data for 1D slices along the ¹³C dimension at indicated positions in 2D NCACX spectrum
• Figure S9a: 2D NCOCX data on 66‒140 fibrils
• Figure S9b: Data for 1D slices along the ¹³C dimension at indicated positions in 2D NCOCX spectrum
• Figure S10a and S10b: 3D NCACX data on labeled 66‒140 fibrils showing representative 2D planes at constant ¹⁵N frequencies
• Figure S10c and S10d: Data for 1D slices along the ¹³C dimension at indicated positions in 3D NCaCX spectrum
• Figure S11a and S11b: 3D NCOCX spectra on labeled 66‒140 fibrils showing representative 2D planes at constant ¹⁵N frequencies
• Figure S11c and S11d: Data for 1D slices along the ¹³C dimension at indicated positions in 3D NCaCX spectrum
• Figure S12a and S12b: 3D NCACX data on labeled 66‒140 fibrils showing representative 2D planes at constant ¹³Ca frequencies
• Figure S12c and S12d: Data for 1D slices along the ¹³C dimension at indicated positions in 3D NCaCX spectrum
• Figure S13a: Data for 1D ¹³C on labeled 66-140 fibrils
• Figure S13b: Data for 1D ¹⁵N on labeled 66-140 fibrils
• Figure S13c: Data for frequency-selective REDOR experiments
• Figure S16: Raman data of single-Ala mutants
• Figure S17: ThT aggregation kinetic data on cross-seeding of soluble 66‒140
• Figure S18: ThT aggregation kinetic data on E104A/E105A seeded with 66‒140 fibrils
• Figure S19a and S19b:ThT aggregation kinetic data on K96A/K97A and cross-seeding by 66‒140 fibrils
• Figure S20:ThT aggregation kinetic data of soluble single-Ala mutants seeded with 66‒140 fibrils
• Figure S25b: 66-140 molecular dynamic simulation data on radial distribution functions for distance of K80, K96 and K97 to E83 in 66-140
• Figure S25d: 66-140 molecular dynamic simulation data for salt bridges occurring over 15-ns
• Figure S26: ThT aggregation kinetic data of Ac1-140 and single Ala-mutants
• Figure S28: Raman data for Ac1–140 and single Ala-mutants

Figures and Tables

Figure 2b and S5a: TEM image of 66-140 fibrils

Figure 2b and S5b: AFM image of 66-140 fibrils

Figure 3c: Structural model of the 66‒140 fibril

Figure 4c and S15: TEM images of K80A, E83A, K96A, K97A, D98A and K102A fibrils

Figure 4c and S15: AFM images of K80A, E83A, K96A, K97A, D98A and K102A fibrils

Figure 6B and S21: TEM images of K80A, E83A, K96A, K97A, D98A and K102A mutants seeded with 66-140 fibrils

Figure 7B and S23: TEM image of E83K/K97E seeded with 66‒140 fibrils

Figure 8F and S27: TEM images of Ac1‒140 and single Ala-mutants

Supplementary Figure 1a: SDS-PAGE gel showing CtsB degradation of soluble syn

Supplementary Figure 1b: SDS-PAGE gel showing CtsL degradation of soluble syn

Supplementary Figure 1c: SDS-PAGE gel showing AEP degradation of soluble syn

Supplementary Figure 1d: SDS-PAGE gel showing CtsD degradation of soluble syn

Supplementary Figure 3: TEM images 66–140 fibrils taken post aggregation in 0.4 and 0.8 M NaCl

Supplementary Figure 4B: TEM images of 66–140 seeded with 66-140 fibrils

Supplementary Figure 25: Data for 66–140 model structure by molecular dynamics simulations

Supplementary Table 6: PK digestion of 66‒140 fibrils

Supplementary Table 7: Reported _tlag and _t½ for 66–140 and single-Ala aggregation kinetic data

Supplementary Table 8: PK digestion of single-Ala mutant fibrils