Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1993 Jan 1;90(1):1–5. doi: 10.1073/pnas.90.1.1

Perturbation of the secondary structure of the scrapie prion protein under conditions that alter infectivity.

M Gasset 1, M A Baldwin 1, R J Fletterick 1, S B Prusiner 1
PMCID: PMC45587  PMID: 8419912

Abstract

Limited proteolysis of the scrapie prion protein (PrPSc) generates PrP 27-30, which polymerizes into amyloid. By attenuated total reflection-Fourier transform infrared spectroscopy, PrP 27-30 polymers contained 54% beta-sheet, 25% alpha-helix, 10% turns, and 11% random coil; dispersion into detergent-lipid-protein-complexes preserved infectivity and secondary structure. Almost 60% of the beta-sheet was low-frequency infrared-absorbing, reflecting intermolecular aggregation. Decreased low-frequency beta-sheet and increased turn content were found after SDS/PAGE, which disassembled the amyloid polymers, denatured PrP 27-30, and diminished scrapie infectivity. Acid-induced transitions were reversible, whereas alkali produced an irreversible transition centered at pH 10 under conditions that diminished infectivity. Whether PrPSc synthesis involves a transition in the secondary structure of one or more domains of the cellular prion protein from alpha-helical, random coil, or turn into beta-sheet remains to be established.

Full text

PDF
3

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bazan J. F., Fletterick R. J., McKinley M. P., Prusiner S. B. Predicted secondary structure and membrane topology of the scrapie prion protein. Protein Eng. 1987 Feb-Mar;1(2):125–135. doi: 10.1093/protein/1.2.125. [DOI] [PubMed] [Google Scholar]
  2. Bolton D. C., Bendheim P. E., Marmorstein A. D., Potempska A. Isolation and structural studies of the intact scrapie agent protein. Arch Biochem Biophys. 1987 Nov 1;258(2):579–590. doi: 10.1016/0003-9861(87)90380-8. [DOI] [PubMed] [Google Scholar]
  3. Bolton D. C., McKinley M. P., Prusiner S. B. Identification of a protein that purifies with the scrapie prion. Science. 1982 Dec 24;218(4579):1309–1311. doi: 10.1126/science.6815801. [DOI] [PubMed] [Google Scholar]
  4. Bolton D. C., McKinley M. P., Prusiner S. B. Molecular characteristics of the major scrapie prion protein. Biochemistry. 1984 Dec 4;23(25):5898–5906. doi: 10.1021/bi00320a002. [DOI] [PubMed] [Google Scholar]
  5. Borchelt D. R., Scott M., Taraboulos A., Stahl N., Prusiner S. B. Scrapie and cellular prion proteins differ in their kinetics of synthesis and topology in cultured cells. J Cell Biol. 1990 Mar;110(3):743–752. doi: 10.1083/jcb.110.3.743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brown P., Rohwer R. G., Gajdusek D. C. Sodium hydroxide decontamination of Creutzfeldt-Jakob disease virus. N Engl J Med. 1984 Mar 15;310(11):727–727. doi: 10.1056/NEJM198403153101122. [DOI] [PubMed] [Google Scholar]
  7. Cabiaux V., Brasseur R., Wattiez R., Falmagne P., Ruysschaert J. M., Goormaghtigh E. Secondary structure of diphtheria toxin and its fragments interacting with acidic liposomes studied by polarized infrared spectroscopy. J Biol Chem. 1989 Mar 25;264(9):4928–4938. [PubMed] [Google Scholar]
  8. Caughey B. W., Dong A., Bhat K. S., Ernst D., Hayes S. F., Caughey W. S. Secondary structure analysis of the scrapie-associated protein PrP 27-30 in water by infrared spectroscopy. Biochemistry. 1991 Aug 6;30(31):7672–7680. doi: 10.1021/bi00245a003. [DOI] [PubMed] [Google Scholar]
  9. Caughey B., Raymond G. J. The scrapie-associated form of PrP is made from a cell surface precursor that is both protease- and phospholipase-sensitive. J Biol Chem. 1991 Sep 25;266(27):18217–18223. [PubMed] [Google Scholar]
  10. Chirgadze Y. N., Fedorov O. V., Trushina N. P. Estimation of amino acid residue side-chain absorption in the infrared spectra of protein solutions in heavy water. Biopolymers. 1975 Apr;14(4):679–694. doi: 10.1002/bip.1975.360140402. [DOI] [PubMed] [Google Scholar]
  11. DeArmond S. J., McKinley M. P., Barry R. A., Braunfeld M. B., McColloch J. R., Prusiner S. B. Identification of prion amyloid filaments in scrapie-infected brain. Cell. 1985 May;41(1):221–235. doi: 10.1016/0092-8674(85)90076-5. [DOI] [PubMed] [Google Scholar]
  12. Diringer H., Gelderblom H., Hilmert H., Ozel M., Edelbluth C., Kimberlin R. H. Scrapie infectivity, fibrils and low molecular weight protein. Nature. 1983 Dec 1;306(5942):476–478. doi: 10.1038/306476a0. [DOI] [PubMed] [Google Scholar]
  13. Endo T., Groth D., Prusiner S. B., Kobata A. Diversity of oligosaccharide structures linked to asparagines of the scrapie prion protein. Biochemistry. 1989 Oct 17;28(21):8380–8388. doi: 10.1021/bi00447a017. [DOI] [PubMed] [Google Scholar]
  14. Fraser P. E., Nguyen J. T., Surewicz W. K., Kirschner D. A. pH-dependent structural transitions of Alzheimer amyloid peptides. Biophys J. 1991 Nov;60(5):1190–1201. doi: 10.1016/S0006-3495(91)82154-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gabizon R., McKinley M. P., Groth D. F., Kenaga L., Prusiner S. B. Properties of scrapie prion protein liposomes. J Biol Chem. 1988 Apr 5;263(10):4950–4955. [PubMed] [Google Scholar]
  16. Gabizon R., McKinley M. P., Prusiner S. B. Purified prion proteins and scrapie infectivity copartition into liposomes. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4017–4021. doi: 10.1073/pnas.84.12.4017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gajdusek D. C. Unconventional viruses and the origin and disappearance of kuru. Science. 1977 Sep 2;197(4307):943–960. doi: 10.1126/science.142303. [DOI] [PubMed] [Google Scholar]
  18. Garnier J., Osguthorpe D. J., Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. doi: 10.1016/0022-2836(78)90297-8. [DOI] [PubMed] [Google Scholar]
  19. Glenner G. G. Amyloid deposits and amyloidosis. The beta-fibrilloses (first of two parts). N Engl J Med. 1980 Jun 5;302(23):1283–1292. doi: 10.1056/NEJM198006053022305. [DOI] [PubMed] [Google Scholar]
  20. Glenner G. G., Eanes E. D., Bladen H. A., Linke R. P., Termine J. D. Beta-pleated sheet fibrils. A comparison of native amyloid with synthetic protein fibrils. J Histochem Cytochem. 1974 Dec;22(12):1141–1158. doi: 10.1177/22.12.1141. [DOI] [PubMed] [Google Scholar]
  21. Goormaghtigh E., Cabiaux V., Ruysschaert J. M. Secondary structure and dosage of soluble and membrane proteins by attenuated total reflection Fourier-transform infrared spectroscopy on hydrated films. Eur J Biochem. 1990 Oct 24;193(2):409–420. doi: 10.1111/j.1432-1033.1990.tb19354.x. [DOI] [PubMed] [Google Scholar]
  22. Hemström C., Virtanen A., Bridge E., Ketner G., Pettersson U. Adenovirus E4-dependent activation of the early E2 promoter is insufficient to promote the early-to-late-phase transition. J Virol. 1991 Mar;65(3):1440–1449. doi: 10.1128/jvi.65.3.1440-1449.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hsiao K., Dlouhy S. R., Farlow M. R., Cass C., Da Costa M., Conneally P. M., Hodes M. E., Ghetti B., Prusiner S. B. Mutant prion proteins in Gerstmann-Sträussler-Scheinker disease with neurofibrillary tangles. Nat Genet. 1992 Apr;1(1):68–71. doi: 10.1038/ng0492-68. [DOI] [PubMed] [Google Scholar]
  24. Jackson M., Mantsch H. H. Beware of proteins in DMSO. Biochim Biophys Acta. 1991 Jun 24;1078(2):231–235. doi: 10.1016/0167-4838(91)90563-f. [DOI] [PubMed] [Google Scholar]
  25. Kascsak R. J., Rubenstein R., Merz P. A., Tonna-DeMasi M., Fersko R., Carp R. I., Wisniewski H. M., Diringer H. Mouse polyclonal and monoclonal antibody to scrapie-associated fibril proteins. J Virol. 1987 Dec;61(12):3688–3693. doi: 10.1128/jvi.61.12.3688-3693.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kennedy D. F., Crisma M., Toniolo C., Chapman D. Studies of peptides forming 3(10)- and alpha-helices and beta-bend ribbon structures in organic solution and in model biomembranes by Fourier transform infrared spectroscopy. Biochemistry. 1991 Jul 2;30(26):6541–6548. doi: 10.1021/bi00240a026. [DOI] [PubMed] [Google Scholar]
  27. McKinley M. P., Braunfeld M. B., Bellinger C. G., Prusiner S. B. Molecular characteristics of prion rods purified from scrapie-infected hamster brains. J Infect Dis. 1986 Jul;154(1):110–120. doi: 10.1093/infdis/154.1.110. [DOI] [PubMed] [Google Scholar]
  28. Merz P. A., Somerville R. A., Wisniewski H. M., Iqbal K. Abnormal fibrils from scrapie-infected brain. Acta Neuropathol. 1981;54(1):63–74. doi: 10.1007/BF00691333. [DOI] [PubMed] [Google Scholar]
  29. Meyer R. K., McKinley M. P., Bowman K. A., Braunfeld M. B., Barry R. A., Prusiner S. B. Separation and properties of cellular and scrapie prion proteins. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2310–2314. doi: 10.1073/pnas.83.8.2310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Prusiner S. B., Bolton D. C., Groth D. F., Bowman K. A., Cochran S. P., McKinley M. P. Further purification and characterization of scrapie prions. Biochemistry. 1982 Dec 21;21(26):6942–6950. doi: 10.1021/bi00269a050. [DOI] [PubMed] [Google Scholar]
  31. Prusiner S. B., Groth D. F., Bolton D. C., Kent S. B., Hood L. E. Purification and structural studies of a major scrapie prion protein. Cell. 1984 Aug;38(1):127–134. doi: 10.1016/0092-8674(84)90533-6. [DOI] [PubMed] [Google Scholar]
  32. Prusiner S. B., Groth D. F., McKinley M. P., Cochran S. P., Bowman K. A., Kasper K. C. Thiocyanate and hydroxyl ions inactivate the scrapie agent. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4606–4610. doi: 10.1073/pnas.78.7.4606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Prusiner S. B., McKinley M. P., Bowman K. A., Bolton D. C., Bendheim P. E., Groth D. F., Glenner G. G. Scrapie prions aggregate to form amyloid-like birefringent rods. Cell. 1983 Dec;35(2 Pt 1):349–358. doi: 10.1016/0092-8674(83)90168-x. [DOI] [PubMed] [Google Scholar]
  34. Prusiner S. B. Molecular biology of prion diseases. Science. 1991 Jun 14;252(5012):1515–1522. doi: 10.1126/science.1675487. [DOI] [PubMed] [Google Scholar]
  35. Saitta B., Timpl R., Chu M. L. Human alpha 2(VI) collagen gene. Heterogeneity at the 5'-untranslated region generated by an alternate exon. J Biol Chem. 1992 Mar 25;267(9):6188–6196. [PubMed] [Google Scholar]
  36. Serban D., Taraboulos A., DeArmond S. J., Prusiner S. B. Rapid detection of Creutzfeldt-Jakob disease and scrapie prion proteins. Neurology. 1990 Jan;40(1):110–117. doi: 10.1212/wnl.40.1.110. [DOI] [PubMed] [Google Scholar]
  37. Stahl N., Baldwin M. A., Burlingame A. L., Prusiner S. B. Identification of glycoinositol phospholipid linked and truncated forms of the scrapie prion protein. Biochemistry. 1990 Sep 25;29(38):8879–8884. doi: 10.1021/bi00490a001. [DOI] [PubMed] [Google Scholar]
  38. Stahl N., Borchelt D. R., Prusiner S. B. Differential release of cellular and scrapie prion proteins from cellular membranes by phosphatidylinositol-specific phospholipase C. Biochemistry. 1990 Jun 5;29(22):5405–5412. doi: 10.1021/bi00474a028. [DOI] [PubMed] [Google Scholar]
  39. Surewicz W. K., Leddy J. J., Mantsch H. H. Structure, stability, and receptor interaction of cholera toxin as studied by Fourier-transform infrared spectroscopy. Biochemistry. 1990 Sep 4;29(35):8106–8111. doi: 10.1021/bi00487a017. [DOI] [PubMed] [Google Scholar]
  40. Tagliavini F., Prelli F., Ghiso J., Bugiani O., Serban D., Prusiner S. B., Farlow M. R., Ghetti B., Frangione B. Amyloid protein of Gerstmann-Sträussler-Scheinker disease (Indiana kindred) is an 11 kd fragment of prion protein with an N-terminal glycine at codon 58. EMBO J. 1991 Mar;10(3):513–519. doi: 10.1002/j.1460-2075.1991.tb07977.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES