DOI: 10.25444/nhlbi.28513553.v1
title: Data associated with "Defining essential charged residues in fibril formation of a lysosomal derived N-terminal alpha-synuclein truncation

description: <b>Source data</b>:Figure 1d-g: Activity assays for CtsB, CtsL, CtsD and AEPFigure 2a: ThT aggregation kinetic data for Ac1-140 and 66-140Figure 2c and S6: Raman data on Ac1-140 and 66-140 fibrilsFigure 3a: 2D <sup>13</sup>C-<sup>13</sup>C ssNMR data for 66-140 fibrilsFigure 3b: Secondary chemical shift data for 66-140Figure 4b and S14: ThT aggregation kinetic data for 66-140 and single ala-mutantsFigure 5a and S16: Raman data 66-140 and single ala-mutantFigure 5b: Raman difference data subtracting 66-140 from single ala-mutantsFigure 5c: ThT aggregation kinetic data of soluble 66-140 seeded with single ala-mutant fibrilsFigure 6a: ThT aggregation kinetic data of single ala-mutants seeded with 66-140 fibrilsFigure 6c and S22: Raman data of single-Ala mutants seeded with 66–140 fibrilsFigure 7a: ThT aggregation kinetic data of charge switch mutants seeded with 66-140 fibrilsFigure 7c and S24: Raman data of E83K/K97E seeded with 66–140 fibrilsFigure 8b: ThT aggregation kinetic data for Ac1-140Figure 8c: ThT aggregation kinetic data AcK80AFigure 8d: ThT aggregation kinetic data AcK96AFigure 8e: ThT aggregation kinetic data AcK97AFigure 8g: ThT aggregation kinetic data of soluble Ac1-140 seeded with single-Ala full-length mutantsFigure S3: Data for a salt-dependent aggregation of 66–140Figure S4a: ThT aggregation kinetic data for self-seeding 66‒140Figure S7a: 2D carbon-carbon cross-polarization dataFigure S7b: Data for 1D slices along the <sup>13</sup>C dimension at indicated positions in 2D carbon-carbon spectrumFigure S8a: 2D NCACX data on 66‒140 fibrilsFigure S8b: Data for 1D slices along the <sup>13</sup>C dimension at indicated positions in 2D NCACX spectrumFigure S9a: 2D NCOCX data on 66‒140 fibrilsFigure S9b: Data for 1D slices along the <sup>13</sup>C dimension at indicated positions in 2D NCOCX spectrumFigure S10a and S10b: 3D NCACX data on labeled 66‒140 fibrils showing representative 2D planes at constant <sup>15</sup>N frequenciesFigure S10c and S10d: Data for 1D slices along the <sup>13</sup>C dimension at indicated positions in 3D NCaCX spectrumFigure S11a and S11b: 3D NCOCX spectra on labeled 66‒140 fibrils showing representative 2D planes at constant <sup>15</sup>N frequenciesFigure S11c and S11d: Data for 1D slices along the <sup>13</sup>C dimension at indicated positions in 3D NCaCX spectrumFigure S12a and S12b: 3D NCACX data on labeled 66‒140 fibrils showing representative 2D planes at constant <sup>13</sup>Ca frequenciesFigure S12c and S12d: Data for 1D slices along the <sup>13</sup>C dimension at indicated positions in 3D NCaCX spectrumFigure S13a: Data for 1D <sup>13</sup>C on labeled 66-140 fibrilsFigure S13b: Data for 1D <sup>15</sup>N on labeled 66-140 fibrilsFigure S13c: Data for frequency-selective REDOR experimentsFigure S16: Raman data of single-Ala mutantsFigure S17: ThT aggregation kinetic data on cross-seeding of soluble 66‒140Figure S18: ThT aggregation kinetic data on E104A/E105A seeded with 66‒140 fibrilsFigure S19a and S19b:ThT aggregation kinetic data on K96A/K97A and cross-seeding by 66‒140 fibrilsFigure S20:ThT aggregation kinetic data of soluble single-Ala mutants seeded with 66‒140 fibrilsFigure S25b: 66-140 molecular dynamic simulation data on radial distribution functions for distance of K80, K96 and K97 to E83 in 66-140Figure S25d: 66-140 molecular dynamic simulation data for salt bridges occurring over 15-nsFigure S26: ThT aggregation kinetic data of Ac1-140 and single Ala-mutantsFigure S28: Raman data for Ac1–140 and single Ala-mutants<b>Figures and Tables</b><br>Figure 2b and S5a: TEM image of 66-140 fibrilsFigure 2b and S5b: AFM image of 66-140 fibrilsFigure 3c: Structural model of the 66‒140 fibrilFigure 4c and S15: TEM images of K80A, E83A, K96A, K97A, D98A and K102A fibrilsFigure 4c and S15: AFM images of K80A, E83A, K96A, K97A, D98A and K102A fibrilsFigure 6B and S21: TEM images of K80A, E83A, K96A, K97A, D98A and K102A mutants seeded with 66-140 fibrilsFigure 7B and S23: TEM image of E83K/K97E seeded with 66‒140 fibrilsFigure 8F and S27: TEM images of Ac1‒140 and single Ala-mutantsSupplementary Figure 1a: SDS-PAGE gel showing CtsB degradation of soluble synSupplementary Figure 1b: SDS-PAGE gel showing CtsL degradation of soluble synSupplementary Figure 1c: SDS-PAGE gel showing AEP degradation of soluble synSupplementary Figure 1d: SDS-PAGE gel showing CtsD degradation of soluble synSupplementary Figure 3: TEM images 66–140 fibrils taken post aggregation in 0.4 and 0.8 M NaClSupplementary Figure 4B: TEM images of 66–140 seeded with 66-140 fibrilsSupplementary Figure 25: Data for 66–140 model structure by molecular dynamics simulationsSupplementary Table 6: PK digestion of 66‒140 fibrilsSupplementary Table 7: Reported <sub>t</sub>lag and <sub>t</sub>½ for 66–140 and single-Ala aggregation kinetic dataSupplementary Table 8: PK digestion of single-Ala mutant fibrils<br><br><br><br>