Journal home
Advance online publication
Current issue
Archive
Press releases
Supplements
Focuses
Guide to authors
Online submissionOnline submission
For referees
Free online issue
Contact the journal
Subscribe
Advertising
work@npg
Reprints and permissions
About this site
For librarians
 
NPG Resources
Nature
Nature Reviews
Nature Immunology
Nature Cell Biology
Nature Genetics
news@nature.com
Nature Conferences
Dissect Medicine
NPG Subject areas
Biotechnology
Cancer
Chemistry
Clinical Medicine
Dentistry
Development
Drug Discovery
Earth Sciences
Evolution & Ecology
Genetics
Immunology
Materials Science
Medical Research
Microbiology
Molecular Cell Biology
Neuroscience
Pharmacology
Physics
Browse all publications
Article
Nature Medicine  2, 59 - 64 (1996)
doi:10.1038/nm0196-59

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

Ruth Gabizon1, Glenn Telling2, Zeev Meiner1, Michele Halimi1, Irit Kahana1 & Stanley B. Prusiner2, 3

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

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

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

 Correspondence should be addressed to R.G.

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.

REFERENCES
  1. Prusiner, S.B. Inherited prion diseases. Proc. Natl. Acad. Sci. USA 91, 4611−4614 (1994).
  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).
  3. Hsiao, K. et al. Linkage of a prion protein missense variant to Gerstmann-Straussler syndrome. Nature 338, 342−345 (1989).
  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).
  5. Kahana, E., Milton, A., Braham, J. & Sofer, D. Creutzfeldt−Jakob disease: Focus among Libyan Jews in Israel. Science 183, 90−91 (1974).
  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).
  7. Hsiao, K. et al. Mutationof the prion proteininLibyan Jews with Creutzfeldt-Jakob disease. N. Engl. J. Med. 324, 1091−1097 (1991).
  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).
  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).
  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).
  11. Collinge, J. et al. Inherited prion disease (PrP lysine 200) in Britain: Two case reports. Br. Med. J. 306, 391−392 (1993).
  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).
  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).
  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).
  15. Prusiner, S.B. Molecular biology of prion diseases. Science 252, 1515−1522 (1991).
  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).
  17. Huang, Z. et al. Proposed three-dimensional structure for the cellular prion protein. Proc. Natl. Acad. Sci. USA 91, 7139−7143 (1994).
  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).
  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).
  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).
  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).
  22. Meyer, R.K. et al. Separation and properties of cellular and scrapie prion proteins. Proc. Natl. Acad. Sci. USA 83, 2310−2314 (1986).
  23. Kascsak, R.J. et al. Mouse polyclonal and monoclonal antibody to scrapie-asso-ciated fibril proteins. J. Virol. 61, 3688−3693 (1987).
  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).
  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).
  26. Hsiao, K. et al. Mutant prion proteins in Gerstmann-Straussler-Scheinker disease with neurofibrillary tangles. Nature Genet. 1, 68−71 (1992).
  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).
  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).
  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).
  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).
  31. Gasset, M. et al. Predicted alpha-helical regions of the prion protein when synthesized as peptides form amyloid. Proc. Natl. Acad. Sci. USA 89, 10940−10944 (1992).
  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).
  33. Forloni, G. et al. Neurotoxicity of a prion protein fragment. Nature 362, 543−546 (1993).
  34. Nguyen, J., Baldwin, M.A., Cohen, F.E. & Prusiner, S.B. Prion protein peptides induce alpha-helix to beta-sheet conformation altransitions. Biochemistry 34, 4186−4192 (1995).
  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).
  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).
  37. Prusiner, S.B. et al. Transgenetic studies implicate interactions between homologous PrP isoforms in scrapie prion replication. Cell 63, 673−686 (1990).
  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).
  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).
  40. Diringer, H. & Ehlers, B. Chemoprophylaxis of scrapie in mice. J. Gen. Virol. 72, 457−460 (1991).
  41. Caughey, B. & Race, R.E. Potent inhibition of scrapie-associated PrP accumulation by Congo red. J. Neumchem. 59, 768−771 (1992).
  42. Caughey, B. & Raymond, G.J. Sulfated polyanion inhibition of scrapie-associated PrP accumulation in cultured cells. J. Virol. 67, 643−650 (1993).
  43. Ladogana, A. et al. Sulphate polyanions prolong the incubation period of scrapie-infected hamsters. J. Gen. Virol. 73, 661−665 (1992).
  44. Ingrosso, L., Ladogana, A. & Pocchiari, M. Congo red prolongs the incubation period in scrapie-infected hamsters. J. Virol. 69, 506−508 (1995).
  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).
  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).
  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).
 Top
 Top
Abstract
Previous | Next
Table of contents
Download PDFDownload PDF
Send to a friendSend to a friend
Save this linkSave this link

Open Innovation Challenges

naturejobs

References
Export citation
Export references
natureproducts

Search buyers guide:

 
ADVERTISEMENT
 
Nature Medicine
ISSN: 1078-8956
EISSN: 1546-170X
Journal home | Advance online publication | Current issue | Archive | Press releases | Supplements | Focuses | For authors | Online submission | For referees | Free online issue | About the journal | Contact the journal | Subscribe | Advertising | work@npg | Reprints and permissions | About this site | For librarians
Nature Publishing Group, publisher of Nature, and other science journals and reference works©1996 Nature Publishing Group | Privacy policy