Science Advances

Supplementary Materials

This PDF file includes:

  • Supplementary Methods
  • Supplementary Results
  • fig. S1. Fluorescence images with different amounts of fluorescent streptavidin on the substrate.
  • fig. S2. Calibration plot of the number of streptavidin versus fluorescence intensity.
  • fig. S3. Histogram of unbinding force for PrP(23–231) measured in the absence of divalent ions.
  • fig. S4. Histogram of unbinding force for PrP(23–231) measured in 1 mM Mn2+.
  • fig. S5. Histogram of unbinding force for PrP(23–231) measured in 1 mM Ni2+.
  • fig. S6. Histogram of unbinding force for PrP(23–231) measured in 1 mM Cu2+.
  • fig. S7. Formation of PrP(23–230) seeds monitored in real time using ThT fluorescence intensity.
  • fig. S8. No seeds were formed when PrP(90–230) was incubated with divalent metal ions for 84 hours.
  • fig. S9. Seeding activity of PrP(23–230) seeds generated in 10 μM Mn2+ measured using RT-QuIC.
  • fig. S10. Seeding activity of PrP(23–230) seeds generated in 10 μM Ni2+measured using RT-QuIC.
  • fig. S11. Seeding activity of PrP(23–230) seeds generated in 10 μM Cu2+ measured using RT-QuIC.
  • fig. S12. Formation of PrP(23–230) seeds using 1 μM Cu2+ and its corresponding seeding activity.
  • fig. S13. Upon addition of protein seeds to brain slice cultures, residual copper does not increase levels of PKC-δand Bax.
  • fig. S14. Biotinylation of PrP and functionalization with PEG tethers do not alter sensitivity to PK digestion.
  • fig. S15. PrP(23–230) and PrP(90–230) remain in a native conformation after reduction of disulfide bond and functionalization with PEG tethers.
  • fig. S16. One-dimensional 1H-NMR spectra of PrP(23–231) show that the protein is in a natively folded conformation.
  • fig. S17. One-dimensional 1H-NMR spectra of PrP(90–231) show that the protein is in a natively folded conformation.
  • fig. S18. One-dimensional 1H-NMR spectra of PrP(23–230) show that the protein is in a natively folded conformation.
  • fig. S19. One-dimensional 1H-NMR spectra of PrP(90–230) show that the protein is in a natively folded conformation.
  • fig. S20. One-dimensional 1H-NMR spectra of PrP(23–230) after disulfide bond reduction show that the protein remains in a natively folded conformation.
  • fig. S21. One-dimensional 1H-NMR spectra of PrP(90–230) after disulfide bond reduction show that the protein remains in a natively folded conformation.
  • fig. S22. One-dimensional 1H-NMR spectra of PrP(23–230) linked to PEG tethers show that the protein remains in a natively folded conformation.
  • fig. S23. One-dimensional 1H-NMR spectra of PrP(90–230) linked to PEG tethers show that the protein remains in a natively folded conformation.
  • fig. S24. Functionalization with PEG tethers does not alter the thermal stabilities of PrP(23–230) and PrP(90–230).
  • fig. S25. Reduction of disulfide bond and functionalization with PEG tethers do not cause aggregation of PrP(23–230) and PrP(90–230).
  • fig. S26. Protein age and the presence of N-terminal His tag do not cause aggregation of PrP(23–230) and PrP(90–230).
  • fig. S27. Protein age and the presence of N-terminal His tag do not alter the secondary structure of PrP(23–230) and PrP(90–230).
  • table S1. Surface density of PrP in PK digestion experiments.
  • table S2. Off-rate, relative on-rate, and relative association constant (KA) for PrP(23–231) binding.

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