Supplementary Materials

Supplementary Material for:

Peptide probes detect misfolded transthyretin oligomers in plasma of hereditary amyloidosis patients

Joseph D. Schonhoft, Cecilia Monteiro, Lars Plate, Yvonne S. Eisele, John M. Kelly, Daniel Boland, Christopher G. Parker, Benjamin F. Cravatt, Sergio Teruya, Stephen Helmke, Mathew Maurer, John Berk, Yoshiki Sekijima, Marta Novais, Teresa Coelho, Evan T. Powers, Jeffery W. Kelly*

*Corresponding author. Email: jkelly{at}scripps.edu

Published 13 September 2017, Sci. Transl. Med. 9, eaam7621 (2017)
DOI: 10.1126/scitranslmed.aam7621

This PDF file includes:

  • Materials and Methods
  • Fig. S1. The B β-strand of TTR labels TTR50–127 oligomers.
  • Fig. S2. The minimal binding motif is VAVHVF.
  • Fig. S3. Determination of probe stoichiometry in MTTR oligomers.
  • Fig. S4. Additional microscopy images of salivary gland biopsies stained with the B-peptide.
  • Fig. S5. Probe B-1 linearly labels MTTR oligomers in human healthy plasma.
  • Fig. S6. A similar probe B-1 structure-activity relationship is found when MTTR oligomers are incubated in healthy plasma.
  • Fig. S7. Labeling of the high-MW SEC fraction in FAP patients by the B-peptide is fluorophore-independent.
  • Fig. S8. Alanine substitutions on probe B-1 have identical effects on incorporation of the peptide into the high-MW fraction of FAP V30M patient plasma.
  • Fig. S9. B-1 is the only peptide of the TTR β-strands that incorporates into the high-MW fraction of patient plasma.
  • Fig. S10. Diazirine-containing probe B-2 selectively labels oligomeric TTR.
  • Fig. S11. Schematic of probe B-2 non-native TTR gel quantification method and representative data.
  • Fig. S12. Probe B-1 does not cross-react with the anti-TTR antibody (DAKO, catalog no. A0002).
  • Fig. S13. Correlation of spectral counts in the MS1 spectra of the diazirine-containing B-2 targets from V30M FAP patients (average of three patients) with plasma concentration.
  • Fig. S14. Validation of N-terminally cleaved non-native TTR as a target of the B-peptide in V30M FAP patient plasma.

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Other Supplementary Material for this manuscript includes the following:

  • Table S1. Full summary of MudPIT LC-MS/MS data presented in Fig. 5 (Excel format).
  • Table S2. All raw data for experiments where n < 20 (Excel format).

[Download Tables S1 and S2]