Neurochemical Research

, Volume 44, Issue 6, pp 1399–1409 | Cite as

Segments in the Amyloid Core that Distinguish Hamster from Mouse Prion Fibrils

  • Howard C.-H. Shen
  • Yung-Han Chen
  • Yu-Sheng Lin
  • Brett K.-Y. Chu
  • Ching-Shin Liang
  • Chien-Chih Yang
  • Rita P.-Y. ChenEmail author
Original Paper


Prion diseases are transmissible fatal neurodegenerative disorders affecting humans and other mammals. The disease transmission can occur between different species but is limited by the sequence homology between host and inoculum. The crucial molecular event in the progression of this disease is prion formation, starting from the conformational conversion of the normal, membrane-anchored prion protein (PrPC) into the misfolded, β-sheet-rich and aggregation-prone isoform (PrPSc), which then self-associates into the infectious amyloid form called prion. Amyloid is the aggregate formed from one-dimensional protein association. As amyloid formation is a key hallmark in prion pathogenesis, studying which segments in prion protein are involved in the amyloid formation can provide molecular details in the cross-species transmission barrier of prion diseases. However, due to the difficulties of studying protein aggregates, very limited knowledge about prion structure or prion formation was disclosed by now. In this study, cross-seeding assay was used to identify the segments involved in the amyloid fibril formation of full-length hamster prion protein, SHaPrP(23–231). Our results showed that the residues in the segments 108–127, 172–194 (helix 2 in PrPC) and 200–227 (helix 3 in PrPC) are in the amyloid core of hamster prion fibrils. The segment 127–143, but not 107–126 (which corresponds to hamster sequence 108–127), was previously reported to be involved in the amyloid core of full-length mouse prion fibrils. Our results indicate that hamster prion protein and mouse prion protein use different segments to form the amyloid core in amyloidogenesis. The sequence-dependent core formation can be used to explain the seeding barrier between mouse and hamster.


Prion Amyloid Fibril Hamster Cross-β Seeding 



The TEM images were obtained with the assistance of Mr. Tai-Lang Lin from the core facility of the Institute of Cellular and Organismic Biology, Academia Sinica. Mass spectra were acquired from three mass spectrometry facilities in Academia Sinica. Protein identification by ESI-TOF was conducted in the mass spectrometry facility of the Institute of Chemistry. Peptide identification by MALDI-TOF was conducted in the mass spectrometry facility of the Institute of Molecular Biology. The MS/MS study by LTQ Orbitrap was performed in the Academia Sinica Common Mass Spectrometry Facility for proteomics and protein modification analysis located at the Institute of Biological Chemistry, supported by Academia Sinica Core Facility and Innovative Instrument Project (AS-CFII-108-107). This work was funded by the Ministry of Science and Technology (MOST) of Taiwan (MOST 105-2119-M-001-028).

Compliance with Ethical Standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

11064_2018_2709_MOESM1_ESM.pdf (1 mb)
Supplementary material 1 (PDF 1031 KB)


  1. 1.
    Prusiner SB (1998) Prions. Proc Natl Acad Sci USA 95:13363–13383CrossRefGoogle Scholar
  2. 2.
    Donne DG, Viles JH, Groth D, Mehlhorn I, James TL, Cohen FE, Prusiner SB, Wright PE, Dyson HJ (1997) Structure of the recombinant full-length hamster prion protein PrP(29–231): the N terminus is highly flexible. Proc Natl Acad Sci USA 94:13452–13457CrossRefGoogle Scholar
  3. 3.
    Riek R, Hornemann S, Wider G, Glockshuber R, Wuthrich K (1997) NMR characterization of the full-length recombinant murine prion protein, mPrP(23–231). FEBS Lett 413:282–288CrossRefGoogle Scholar
  4. 4.
    Riek R, Hornemann S, Wider G, Billeter M, Glockshuber R, Wuthrich K (1996) NMR structure of the mouse prion protein domain PrP(121–231). Nature 382:180–182CrossRefGoogle Scholar
  5. 5.
    Pan K, Baldwin M, Nguyen J, Gasset M, Serban A, Groth D, Mehlhorn I, Huang Z, Fletterick R, Cohen F, Prusiner S (1993) Conversion of a-helices b-sheets features in the formation of the scrapie prion proteins. Proc Natl Acad Sci USA 90:10962–10966CrossRefGoogle Scholar
  6. 6.
    Safar J, Roller P, Gajdusek D, Gibbs C Jr (1993) Conformational transitions, dissociation, and unfolding of scrapie amyloid (prion) protein. J Biol Chem 268:20276–20284Google Scholar
  7. 7.
    Lu X, Wintrode PL, Surewicz WK (2007) b-Sheet core of human prion protein amyloid fibrils as determined by hydrogen/deuterium exchange. Proc Natl Acad Sci USA 104:1510–1515CrossRefGoogle Scholar
  8. 8.
    Singh J, Udgaonkar JB (2013) Dissection of conformational conversion events during prion amyloid fibril formation using hydrogen exchange and mass spectrometry. J Mol Biol 425:3510–3521CrossRefGoogle Scholar
  9. 9.
    Nazabal A, Hornemann S, Aguzzi A, Zenobi R (2009) Hydrogen/deuterium exchange mass spectrometry identifies two highly protected regions in recombinant full-length prion protein amyloid fibrils. J Mass Spectrom 44:965–977CrossRefGoogle Scholar
  10. 10.
    Smirnovas V, Baron GS, Offerdahl DK, Raymond GJ, Caughey B, Surewicz WK (2011) Structural organization of brain-derived mammalian prions examined by hydrogen-deuterium exchange. Nat Struct Mol Biol 18:504–506CrossRefGoogle Scholar
  11. 11.
    Smirnovas V, Kim JI, Lu X, Atarashi R, Caughey B, Surewicz WK (2009) Distinct structures of scrapie prion protein (PrPSc)-seeded versus spontaneous recombinant prion protein fibrils revealed by hydrogen/deuterium exchange. J Biol Chem 284:24233–24241CrossRefGoogle Scholar
  12. 12.
    Kuwata K, Matumoto T, Cheng H, Nagayama K, James TL, Roder H (2003) NMR-detected hydrogen exchange and molecular dynamics simulations provide structural insight into fibril formation of prion protein fragment 106–126. Proc Natl Acad Sci USA 100:14790–14795CrossRefGoogle Scholar
  13. 13.
    Cobb NJ, Sonnichsen FD, McHaourab H, Surewicz WK (2007) Molecular architecture of human prion protein amyloid: a parallel, in-register b-structure. Proc Natl Acad Sci USA 104:18946–18951CrossRefGoogle Scholar
  14. 14.
    Helmus JJ, Surewicz K, Apostol MI, Surewicz WK, Jaroniec CP (2011) Intermolecular alignment in Y145Stop human prion protein amyloid fibrils probed by solid-state NMR spectroscopy. J Am Chem Soc 133:13934–13937CrossRefGoogle Scholar
  15. 15.
    Helmus JJ, Surewicz K, Nadaud PS, Surewicz WK, Jaroniec CP (2008) Molecular conformation and dynamics of the Y145Stop variant of human prion protein in amyloid fibrils. Proc Natl Acad Sci USA 105:6284–6289CrossRefGoogle Scholar
  16. 16.
    Helmus JJ, Surewicz K, Surewicz WK, Jaroniec CP (2010) Conformational flexibility of Y145Stop human prion protein amyloid fibrils probed by solid-state nuclear magnetic resonance spectroscopy. J Am Chem Soc 132:2393–2403CrossRefGoogle Scholar
  17. 17.
    Tycko R, Savtchenko R, Ostapchenko VG, Makarava N, Baskakov IV (2010) The alpha-helical C-terminal domain of full-length recombinant PrP converts to an in-register parallel beta-sheet structure in PrP fibrils: evidence from solid state nuclear magnetic resonance. Biochemistry 49:9488–9497CrossRefGoogle Scholar
  18. 18.
    Cheng HM, Tsai TW, Huang WY, Lee HK, Lian HY, Chou FC, Mou Y, Chan JC (2011) Steric zipper formed by hydrophobic peptide fragment of Syrian hamster prion protein. Biochemistry 50:6815–6823CrossRefGoogle Scholar
  19. 19.
    Lin NS, Chao JC, Cheng HM, Chou FC, Chang CF, Chen YR, Chang YJ, Huang SJ, Chan JC (2010) Molecular structure of amyloid fibrils formed by residues 127 to 147 of the human prion protein. Chemistry 16:5492–5499CrossRefGoogle Scholar
  20. 20.
    Lee SW, Mou Y, Lin SY, Chou FC, Tseng WH, Chen CH, Lu CY, Yu SS, Chan JC (2008) Steric zipper of the amyloid fibrils formed by residues 109–122 of the Syrian hamster prion protein. J Mol Biol 378:1142–1154CrossRefGoogle Scholar
  21. 21.
    Vazquez-Fernandez E, Vos MR, Afanasyev P, Cebey L, Sevillano AM, Vidal E, Rosa I, Renault L, Ramos A, Peters PJ, Fernandez JJ, van Heel M, Young HS, Requena JR, Wille H (2016) The structural architecture of an infectious mammalian prion using electron cryomicroscopy. PLoS Pathog 12:e1005835CrossRefGoogle Scholar
  22. 22.
    Watzlawik J, Skora L, Frense D, Griesinger C, Zweckstetter M, Schulz-Schaeffer WJ, Kramer ML (2006) Prion protein helix1 promotes aggregation but is not converted into beta-sheet. J Biol Chem 281:30242–30250Google Scholar
  23. 23.
    Chatterjee B, Lee CY, Lin C, Chen EH, Huang CL, Yang CC, Chen RP (2013) Amyloid core formed of full-length recombinant mouse prion protein involves sequence 127–143 but not sequence 107–126. PLoS ONE 8:e67967CrossRefGoogle Scholar
  24. 24.
    Muramoto T, Scott M, Cohen FE, Prusiner SB (1996) Recombinant scrapie-like prion protein of 106 amino acids is soluble. Proc Natl Acad Sci USA 93:15457–15462CrossRefGoogle Scholar
  25. 25.
    Shindoh R, Kim CL, Song CH, Hasebe R, Horiuchi M (2009) The region approximately between amino acids 81 and 137 of proteinase K-resistant PrPSc is critical for the infectivity of the Chandler prion strain. J Virol 83:3852–3860CrossRefGoogle Scholar
  26. 26.
    Chabry J, Caughey B, Chesebro B (1998) Specific inhibition of in vitro formation of protease-resistant prion protein by synthetic peptides. J Biol Chem 273:13203–13207CrossRefGoogle Scholar
  27. 27.
    Chabry J, Priola SA, Wehrly K, Nishio J, Hope J, Chesebro B (1999) Species-independent inhibition of abnormal prion protein (PrP) formation by a peptide containing a conserved PrP sequence. J Virol 73:6245–6250Google Scholar
  28. 28.
    DeMarco ML, Daggett V (2004) From conversion to aggregation: protofibril formation of the prion protein. Proc Natl Acad Sci USA 101:2293–2298CrossRefGoogle Scholar
  29. 29.
    DeMarco ML, Silveira J, Caughey B, Daggett V (2006) Structural properties of prion protein protofibrils and fibrils: an experimental assessment of atomic models. Biochemistry 45:15573–15582CrossRefGoogle Scholar
  30. 30.
    Chen PY, Lin CC, Chang YT, Lin SC, Chan SI (2002) One O-linked sugar can affect the coil-to-b structural transition of the prion peptide. Proc Natl Acad Sci USA 99:12633–12638CrossRefGoogle Scholar
  31. 31.
    Liao Y-D, Jeng J-C, Wang C-F, Wang S-C, Chang S-T (2004) Removal of N-terminal methionine from recombinant proteins by engineered E. coli methionine aminopeptidase. Prot Sci 13:1802–1810CrossRefGoogle Scholar
  32. 32.
    Dalboge H, Bayne S, Pedersen J (1990) In vivo processing of N-terminal methionine in E. coli. FEBS Lett 266:1–3CrossRefGoogle Scholar
  33. 33.
    Sang JC, Lee CY, Luh FY, Huang YW, Chiang YW, Chen RP (2012) Slow spontaneous α-to-β structural conversion in a non-denaturing neutral condition reveals the intrinsically disordered property of the disulfide-reduced recombinant mouse prion protein. Prion 6:489–497CrossRefGoogle Scholar
  34. 34.
    Krebs MRH, Bromley EHC, Donald AM (2005) The binding of thioflavin-T to amyloid fibrils: localisation and implications. J Struct Biol 149:30–37CrossRefGoogle Scholar
  35. 35.
    Khurana R, Coleman C, Ionescu-Zanetti C, Carter SA, Krishna V, Grover RK, Roy R, Singh S (2005) Mechanism of thioflavin T binding to amyloid fibrils. J Struct Biol 151:229–238CrossRefGoogle Scholar
  36. 36.
    Hsu JC, Chen HL, Snoeberger RC, Luh FY, Lim TS, Hsu C, Chen RP (2013) Thioflavin T and its photo-irradiative derivatives: Exploring their spectroscopic properties in the absence and presence of amyloid fibrils. J Phys Chem B 117:3459–3468CrossRefGoogle Scholar
  37. 37.
    Lee LY, Chen RP (2007) Quantifying the sequence-dependent species barrier between hamster and mouse prions. J Am Chem Soc 129:1644–1652CrossRefGoogle Scholar
  38. 38.
    Singh J, Kumar H, Sabareesan AT, Udgaonkar JB (2014) Rational stabilization of helix 2 of the prion protein prevents its misfolding and oligomerization. J Am Chem Soc 136:16704–16707CrossRefGoogle Scholar
  39. 39.
    Yamaguchi K, Matsumoto T, Kuwata K (2008) Critical region for amyloid fibril formation of mouse prion protein: unusual amyloidogenic properties of the helix 2 peptide. Biochemistry 47:13242–13251CrossRefGoogle Scholar
  40. 40.
    Priola SA, Chesebro B (1995) A single hamster PrP amino acid blocks conversion to protease-resistant PrP in scrapie-infected mouse neuroblastoma cells. J Virol 69:7754–7758Google Scholar
  41. 41.
    Vanik DL, Surewicz KA, Surewicz WK (2004) Molecular basis of barriers for interspecies transmissibility of mammalian prions. Mol Cell 14:139–145CrossRefGoogle Scholar
  42. 42.
    Jones EM, Surewicz WK (2005) Fibril conformation as the basis of species- and strain-dependent seeding specificity of mammalian prion amyloids. Cell 121:63–72CrossRefGoogle Scholar
  43. 43.
    Theint T, Nadaud PS, Aucoin D, Helmus JJ, Pondaven SP, Surewicz K, Surewicz WK, Jaroniec CP (2017) Species-dependent structural polymorphism of Y145Stop prion protein amyloid revealed by solid-state NMR spectroscopy. Nat Commun 8:753CrossRefGoogle Scholar
  44. 44.
    Groveman BR, Dolan MA, Taubner LM, Kraus A, Wickner RB, Caughey B (2014) Parallel in-register intermolecular beta-sheet architectures for prion-seeded prion protein (PrP) amyloids. J Biol Chem 289:24129–24142CrossRefGoogle Scholar
  45. 45.
    Wille H, Bian W, McDonald M, Kendall A, Colby DW, Bloch L, Ollesch J, Borovinskiy AL, Cohen FE, Prusiner SB, Stubbs G (2009) Natural and synthetic prion structure from X-ray fiber diffraction. Proc Natl Acad Sci USA 106:16990–16995CrossRefGoogle Scholar
  46. 46.
    Silva CJ, Vazquez-Fernandez E, Onisko B, Requena JR (2015) Proteinase K and the structure of PrPSc: the good, the bad and the ugly. Virus Res 207:120–126CrossRefGoogle Scholar
  47. 47.
    Wille H, Requena JR (2018) The Structure of PrP(Sc) Prions. Pathogens 7Google Scholar
  48. 48.
    Govaerts C, Wille H, Prusiner SB, Cohen FE (2004) Evidence for assembly of prions with left-handed β-helices into trimers. Proc Natl Acad Sci USA 101:8342–8347CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Biological ChemistryAcademia SinicaTaipeiTaiwan
  2. 2.Institute of Biochemical SciencesNational Taiwan UniversityTaipeiTaiwan
  3. 3.Department of Biochemical Science and TechnologyNational Taiwan UniversityTaipeiTaiwan
  4. 4.Department of ChemistryNational Taiwan UniversityTaipeiTaiwan

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