Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Insoluble wild–type and protease–resistant mutant prion protein in brains of patients with inherited prion disease

Nature Medicine volume 2, pages 59–64 (1996)Cite this article

Abstract

We studied prion proteins (PrP) in skin and brains of Libyan Jews carrying the E200K mutation who died of familial Creutzfeldt–Jakob disease (CJD). Unexpectedly, studies with brain showed that PrP molecules encoded both by the wild–type (wt) and mutant alleles exhibit altered properties characteristic of the prion protein associated with prion diseases (PrPSc). Using monospecific antisera, we found that wtPrP was insoluble in the brains of three patients who were heterozygous for the E200K mutation, whereas mutant PrP was both insoluble and protease–resistant. Our results argue that both wild–type and mutant PrP undergo conformational changes and are particularly intriguing, because the normal isoform PrPc is soluble in nondenaturing detergents and is readily digested by proteases, whereas PrPSc is insoluble and resistant to proteolytic digestion. Our findings indicate that insoluble wtPrP represents a conformational intermediate, the first to be identified, within a pathway in which PrPc is converted to PrPSc.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Prusiner, S.B. Inherited prion diseases. Proc. Natl. Acad. Sci. USA 91, 4611–4614 (1994).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Masters, C.L., Gajdusek, D.C. & Gibbs, C.J., Creutzfeldt-Jakob disease virus isolations from the Gerstmann-Straussler syndrome. Brain 1O4, 559–588 (1981).

    Article  Google Scholar 

  3. Hsiao, K. et al. Linkage of a prion protein missense variant to Gerstmann-Straussler syndrome. Nature 338, 342–345 (1989).

    Article  CAS  PubMed  Google Scholar 

  4. Goldfarb, L.G. et al. Fatal familial insomnia and familial Creutzfeldt-Jakob disease: Disease phenotype determined by a DNA polymorphism. Science 258, 806–808 (1992).

    Article  CAS  PubMed  Google Scholar 

  5. Kahana, E., Milton, A., Braham, J. & Sofer, D. Creutzfeldt–Jakob disease: Focus among Libyan Jews in Israel. Science 183, 90–91 (1974).

    Article  CAS  PubMed  Google Scholar 

  6. Goldfarb, L., Korczyn, A., Brown, P., Chapman, J. & Gajdusek, D.C. Mutation in codon 200 of scrapie amyloid precursor gene linked to Creutzfeldt-Jakob disease in Sephardicjews of Libyan and non-Libyan origin. Lancet 336, 637–638 (1990).

    Article  CAS  PubMed  Google Scholar 

  7. Hsiao, K. et al. Mutationof the prion proteininLibyan Jews with Creutzfeldt-Jakob disease. N. Engl. J. Med. 324, 1091–1097 (1991).

    Article  CAS  PubMed  Google Scholar 

  8. Gabizon, R. et al. Mutation and polymorphism of the prion protein gene in Libyan Jews with Creutzfeldt-Jakob disease. Am. J. Hum. Genet. 33, 828–835 (1993).

    Google Scholar 

  9. Goldfarb, L.G., Mitrova, E., Brown, P., Toh, B.H. & Gajdusek, D.C. Mutation in codon 200 of scrapie amyloid protein gene in two clusters of Creutzfeldt-Jakob disease in Slovakia. Lancet 336, 514–515 (1990).

    Article  CAS  PubMed  Google Scholar 

  10. Brown, P. et al. Familial Creutzfeldt-Jakob disease in Chile is associated with the codon 200 mutation of the PRNP amyloid precursor gene on chromosome 20. J. Neural. Sci. 112, 65–67 (1992).

    Article  CAS  Google Scholar 

  11. Collinge, J. et al. Inherited prion disease (PrP lysine 200) in Britain: Two case reports. Br. Med. J. 306, 391–392 (1993).

    Article  Google Scholar 

  12. Bertoni, J.M., Brown, P., Goldfarb, L., Gajdusek, D. & Omaha, N.E., Creutzfeldt–Jakob disease with the PRNP codon 200lys mutation and supranuclear palsy but without myoclonus or periodic EEG complexes (Abstr). Neurology 42 (No.4 Suppl. 3), 350 (1992).

    Google Scholar 

  13. Chapman, J., Ben-Israel, J., Goldhammer, Y. & Korczyn, A.D. The risk of developing Creutzfeldt–Jakob disease in subjects with the PRNP gene codon 200 point mutation. Neurology 44, 1683–1686 (1994).

    Article  CAS  PubMed  Google Scholar 

  14. Spudich, S. et al. Complete penetrance of Creutzfeldt-Jakob disease in Libyan Jews carrying the E200K mutation in the prion protein gene. Mol. Med. 1, 607–613 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Prusiner, S.B. Molecular biology of prion diseases. Science 252, 1515–1522 (1991).

    Article  CAS  PubMed  Google Scholar 

  16. Pan, K.-M. et al. Conversion of a-helices into b-sheets features in the formation of the scrapie prion proteins. Proc. Natl. Acad. Sci. USA 90, 10962–10966 (1993).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Huang, Z. et al. Proposed three-dimensional structure for the cellular prion protein. Proc. Natl. Acad. Sci. USA 91, 7139–7143 (1994).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Safar, J., Roller, P.P., Gajdusek, D.C. & Gibbs, C.J., Jr. Conformational transitions, dissociation, and unfolding of scrapie amyloid (prion) protein. J. Biol. Chem. 268, 20276–20284 (1993).

    CAS  PubMed  Google Scholar 

  19. Kitamoto, T., Yamaguchi, K., Doh-ura, K. & Tateishi, J. A prion protein missense variant is integrated in kuru plaque cores in patients with Gerstmann-Straussler syndrome. Neurology 41, 306–310 (1991).

    Article  CAS  PubMed  Google Scholar 

  20. Kitamoto, T., Iizuka, R., & Tateishi, J. An amber mutation of prion protein in Gerstmann-Straussler syndrome with mutant PrP plaques. Biochem. Biophys. Res. Commun. 192, 525–531 (1993).

    Article  CAS  PubMed  Google Scholar 

  21. Tagliavini, F. et al. Amyloid fibrils in Gerstmann-Straussler-Scheinker disease (Indiana and Swedish kindreds) express only PrP peptides encoded by the mutant allele. Cell 79, 695–703 (1994).

    Article  CAS  PubMed  Google Scholar 

  22. Meyer, R.K. et al. Separation and properties of cellular and scrapie prion proteins. Proc. Natl. Acad. Sci. USA 83, 2310–2314 (1986).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Kascsak, R.J. et al. Mouse polyclonal and monoclonal antibody to scrapie-asso-ciated fibril proteins. J. Virol. 61, 3688–3693 (1987).

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Rogers, M., Taraboulos, A., Scott, M., Groth, D. & Prusiner, S.B. Intracellular accumulation of the cellular prion protein after mutagenesis of its Asn-linked gly-cosylation sites. Glycobiology 1, 101–109 (1990).

    Article  CAS  PubMed  Google Scholar 

  25. Goldfarb, L.G. et al. New mutation in scrapie amyloid precursor gene (at codon 178) in Finnish Creutzfeldt–Jakob kindred. Lancet 337, 425 (1991).

    Article  CAS  PubMed  Google Scholar 

  26. Hsiao, K. et al. Mutant prion proteins in Gerstmann-Straussler-Scheinker disease with neurofibrillary tangles. Nature Genet. 1, 68–71 (1992).

    Article  CAS  PubMed  Google Scholar 

  27. Monari, L. et al. Fatal familial insomnia and familial Creutzfeldt–Jakob disease: Different prion proteins determined by a DNA polymorphism. Proc. Natl. Acad. Sci. USA 91, 2839–2842 (1994).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Gasset, M., Baldwin, M.A., Fletterick, R.J. & Prusiner, S.B. Perturbation of the secondary structure of the scrapie prion protein under conditions associated with changes in infectivity. Proc. Natl. Acad. Sci. USA 90, 1–5 (1993).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Gabizon, R., McKinley, M.P., Groth, D.F., Kenaga, L. & Prusiner, S.B. Properties of scrapie prion liposomes. J. Biol. Chem. 263, 4950–4955 (1988).

    CAS  PubMed  Google Scholar 

  30. Goldfarb, L.G. et al. Synthetic peptides corresponding to different mutated regions of the amyloid gene in familial Creutzfeldt–Jakob disease show enhanced in vitro formation of morphologically different amyloid fibrils. Proc. Natl. Acad. Sci. USA 90, 4451–4454 (1993).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Gasset, M. et al. Predicted α-helical regions of the prion protein when synthesized as peptides form amyloid. Proc. Natl. Acad. Sci. USA 89, 10940–10944 (1992).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Come, J.H., Fraser, P.E. & Lansbury, P.T., A kinetic model for amyloid formation in the prion diseases: Importance of seeding. Proc. Natl. Acad. Sci. USA 90, 5959–5963 (1993).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Forloni, G. et al. Neurotoxicity of a prion protein fragment. Nature 362, 543–546 (1993).

    Article  CAS  PubMed  Google Scholar 

  34. Nguyen, J., Baldwin, M.A., Cohen, F.E. & Prusiner, S.B. Prion protein peptides induce α-helix to β-sheet conformation altransitions. Biochemistry 34, 4186–4192 (1995).

    Article  CAS  PubMed  Google Scholar 

  35. Meiner, Z., Halimi, M., Polakiewicz, R.D., Prusiner, S.B. & Gabizon, R. Presence of the prion protein in peripheral tissues of Libyan Jews with Creutzfeldt–Jakob disease. Neurology 42, 1355–1360 (1992).

    Article  CAS  PubMed  Google Scholar 

  36. Taraboulos, A. et al. Acquisition of protease resistance by prion proteins in scrapie-infected cells does not require asparagine-linked glycosylation. Proc. Natl. Acad. Sci. USA 87, 8262–8266 (1990).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Prusiner, S.B. et al. Transgenetic studies implicate interactions between homologous PrP isoforms in scrapie prion replication. Cell 63, 673–686 (1990).

    Article  CAS  PubMed  Google Scholar 

  38. Kimberlin, R.H. & Walker, C.A. The antiviral compound HPA-23 can prevent scrapie when administered at the time of infection. Arch. Virol. 78, 9–18 (1983).

    Article  CAS  PubMed  Google Scholar 

  39. Ehlers, B. & Diringer, H. Dextran sulphate 500 delays and prevents mouse scrapie by impairment of agent replication in spleen. J. Gen. Virol. 65, 1325–1330 (1984).

    Article  CAS  PubMed  Google Scholar 

  40. Diringer, H. & Ehlers, B. Chemoprophylaxis of scrapie in mice. J. Gen. Virol. 72, 457–460 (1991).

    Article  CAS  PubMed  Google Scholar 

  41. Caughey, B. & Race, R.E. Potent inhibition of scrapie-associated PrP accumulation by Congo red. J. Neumchem. 59, 768–771 (1992).

    Article  CAS  Google Scholar 

  42. Caughey, B. & Raymond, G.J. Sulfated polyanion inhibition of scrapie-associated PrP accumulation in cultured cells. J. Virol. 67, 643–650 (1993).

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Ladogana, A. et al. Sulphate polyanions prolong the incubation period of scrapie-infected hamsters. J. Gen. Virol. 73, 661–665 (1992).

    Article  CAS  PubMed  Google Scholar 

  44. Ingrosso, L., Ladogana, A. & Pocchiari, M. Congo red prolongs the incubation period in scrapie-infected hamsters. J. Virol. 69, 506–508 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Gabizon, R., Meiner, Z., Halimi, M. & Ben-Sasson, S.A. Heparin-like molecules bind differentially to prion-proteins and change their intracellular metabolic fate. J. Cell. Physiol. 157, (1993).

    Article  CAS  PubMed  Google Scholar 

  46. Telling, G.C. et al. Transmission of Creutzfeldt–Jakob disease from humans to transgenic mice expressing chimeric human-mouse prion protein. Proc. Natl. Acad. Sci. USA 91, 9936–9940 (1994).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Telling, G.C. et al. Prion propagation in mice expressing human and chimeric PrP transgenes implicates the interaction of cellular PrP with another protein. Cell 83, 79–90 (1995).

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neurology, Hadassah University Hospital, Ein Karem, Jerusalem, 91120, Israel

    Ruth Gabizon, Zeev Meiner, Michele Halimi & Irit Kahana

  2. Department of Neurology, HSE-781, University of California, San Francisco, California, 94143, USA

    Glenn Telling & Stanley B. Prusiner

  3. Department of Biochemistry and Biophysics, University of California, San Francisco, California, 94143, USA

    Stanley B. Prusiner

Authors
  1. Ruth Gabizon
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Glenn Telling
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zeev Meiner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Michele Halimi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Irit Kahana
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Stanley B. Prusiner
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gabizon, R., Telling, G., Meiner, Z. et al. Insoluble wild–type and protease–resistant mutant prion protein in brains of patients with inherited prion disease. Nat Med 2, 59–64 (1996). https://doi.org/10.1038/nm0196-59

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm0196-59

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing