1887

Abstract

An increasing number of scrapie cases with atypical characteristics, designated Nor98, have recently been recognized. Here, the proteinase K (PK)-resistant prion protein (PrP) fragments from two Swedish cases of Nor98 atypical scrapie have been characterized. The prominent, fast-migrating band in the distinct Nor98 Western immunoblot electrophoretic profile was determined to be of 7 kDa in size and was accordingly designated Nor98-PrP7. The antigenic composition of Nor98-PrP7, as assayed by a panel of anti-PrP antibodies, revealed that this fragment comprised a mid-region of PrP from around aa 85 to 148. N- and C-terminally truncated fragments spanning the mid-region of PrP have only been observed in the genetic prion disorder Gerstmann–Sträussler–Scheinker disease. It is shown here that the long-term PK resistance of Nor98-PrP7 is reduced compared with that of PrP in classical scrapie. Enzymic deglycosylation did not change the distinct electrophoretic profile of Nor98-PrP7. A previously unidentified, PK-resistant, C-terminal PrP fragment of around 24 kDa was detected and its PK resistance was investigated. After deglycosylation, this fragment migrated as a 14 kDa polypeptide and was designated PrP-CTF14. Antigenic determination and the size of 14 kDa suggested a fragment spanning approximately aa 120–233. The existence of two PK-resistant PrP fragments, Nor98-PrP7 and PrP-CTF14, that share an overlapping region suggests that at least two distinct PrP conformers with different PK-resistant cores are present in brain extracts from Nor98-affected sheep. The structural gene of PrP in three Nor98-affected sheep was analysed, but no mutations were found that could be correlated to the aberrant PK-resistant profile observed.

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2006-06-01
2019-10-19
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References

  1. Benestad, S. L., Sarradin, P., Thu, B., Schonheit, J., Tranulis, M. A. & Bratberg, B. ( 2003; ). Cases of scrapie with unusual features in Norway and designation of a new type, Nor98. Vet Rec 153, 202–208.[CrossRef]
    [Google Scholar]
  2. Brun, A., Castilla, J., Ramírez, M. A. & 8 other authors ( 2004; ). Proteinase K enhanced immunoreactivity of the prion protein-specific monoclonal antibody 2A11. Neurosci Res 48, 75–83.[CrossRef]
    [Google Scholar]
  3. Chen, S. G., Teplow, D. B., Parchi, P., Teller, J. K., Gambetti, P. & Autilio-Gambetti, L. ( 1995; ). Truncated forms of the human prion protein in normal brain and in prion diseases. J Biol Chem 270, 19173–19180.[CrossRef]
    [Google Scholar]
  4. De Bosschere, H., Roels, S., Benestad, S. L. & Vanopdenbosch, E. ( 2004; ). Scrapie case similar to Nor98 diagnosed in Belgium via active surveillance. Vet Rec 155, 707–708.[CrossRef]
    [Google Scholar]
  5. Elvander, M., Engvall, A. & Klingeborn, B. ( 1988; ). Scrapie in sheep in Sweden. Acta Vet Scand 29, 509–510.
    [Google Scholar]
  6. Garssen, G. J., Van Keulen, L. J. M., Farquhar, C. F., Smits, M. A., Jacobs, J. G., Bossers, A., Meloen, R. H. & Langeveld, J. P. ( 2000; ). Applicability of three anti-PrP peptide sera including staining of tonsils and brainstem of sheep with scrapie. Microsc Res Tech 50, 32–39.[CrossRef]
    [Google Scholar]
  7. Gavier-Widen, D., Noremark, M., Benestad, S., Simmons, M., Renström, L., Bratberg, B., Elvander, M. & af Segerstad, C. H. ( 2004; ). Recognition of the Nor98 variant of scrapie in the Swedish sheep population. J Vet Diagn Invest 16, 562–567.[CrossRef]
    [Google Scholar]
  8. Ghetti, B., Piccardo, P., Spillantini, M. G. & 13 other authors ( 1996; ). Vascular variant of prion protein cerebral amyloidosis with τ-positive neurofibrillary tangles: the phenotype of the stop codon 145 mutation in PRNP. Proc Natl Acad Sci U S A 93, 744–748.[CrossRef]
    [Google Scholar]
  9. Goldmann, W., Baylis, M., Chihota, C., Stevenson, E. & Hunter, N. ( 2005; ). Frequencies of PrP gene haplotypes in British sheep flocks and the implications for breeding programmes. J Appl Microbiol 98, 1294–1302.[CrossRef]
    [Google Scholar]
  10. Harmeyer, S., Pfaff, E. & Groschup, M. H. ( 1998; ). Synthetic peptide vaccines yield monoclonal antibodies to cellular and pathological prion proteins of ruminants. J Gen Virol 79, 937–945.
    [Google Scholar]
  11. Hayashi, H. K., Yokoyama, T., Takata, M., Iwamaru, Y., Imamura, M., Ushiki, Y. K. & Shinagawa, M. ( 2005; ). The N-terminal cleavage site of PrPSc from BSE differs from that of PrPSc from scrapie. Biochem Biophys Res Commun 328, 1024–1027.[CrossRef]
    [Google Scholar]
  12. Hope, J., Wood, S. C. E. R., Birkett, C. R., Chong, A., Bruce, M. E., Cairns, D., Goldmann, W., Hunter, N. & Bostock, C. J. ( 1999; ). Molecular analysis of ovine prion protein identifies similarities between BSE and an experimental isolate of natural scrapie, CH1641. J Gen Virol 80, 1–4.
    [Google Scholar]
  13. Kang, S.-C., Li, R., Wang, C., Pan, T., Liu, T., Rubenstein, R., Barnard, G., Wong, B.-S. & Sy, M.-S. ( 2003; ). Guanidine hydrochloride extraction and detection of prion proteins in mouse and hamster prion diseases by ELISA. J Pathol 199, 534–541.[CrossRef]
    [Google Scholar]
  14. Korth, C., Stierli, B., Streit, P. & 14 other authors ( 1997; ). Prion (PrPSc)-specific epitope defined by a monoclonal antibody. Nature 390, 74–77.[CrossRef]
    [Google Scholar]
  15. Li, R., Liu, T., Wong, B.-S. & 7 other authors ( 2000; ). Identification of an epitope in the C terminus of normal prion protein whose expression is modulated by binding events in the N terminus. J Mol Biol 301, 567–573.[CrossRef]
    [Google Scholar]
  16. Monari, L., Chen, S. G., Brown, P. & 12 other authors ( 1994; ). Fatal familial insomnia and familial Creutzfeldt–Jakob disease: different prion proteins determined by a DNA polymorphism. Proc Natl Acad Sci U S A 91, 2839–2842.[CrossRef]
    [Google Scholar]
  17. Montagna, P., Gambetti, P., Cortelli, P. & Lugaresi, E. ( 2003; ). Familial and sporadic fatal insomnia. Lancet Neurol 2, 167–176.[CrossRef]
    [Google Scholar]
  18. Moum, T., Olsaker, I., Hopp, P., Moldal, T., Valheim, M., Moum, T. & Benestad, S. L. ( 2005; ). Polymorphisms at codons 141 and 154 in the ovine prion protein gene are associated with scrapie Nor98 cases. J Gen Virol 86, 231–235.[CrossRef]
    [Google Scholar]
  19. Nelson, R., Sawaya, M. R., Balbirnie, M., Madsen, A. Ø., Riekel, C., Grothe, R. & Eisenberg, D. ( 2005; ). Structure of the cross-β spine of amyloid-like fibrils. Nature 435, 773–778.[CrossRef]
    [Google Scholar]
  20. Onnasch, H., Gunn, H. M., Bradshaw, B. J., Benestad, S. L. & Bassett, H. F. ( 2004; ). Two Irish cases of scrapie resembling Nor98. Vet Rec 155, 636–637.[CrossRef]
    [Google Scholar]
  21. O'Rourke, K. I., Baszler, T. V., Miller, J. M., Spraker, T. R., Sadler-Riggleman, I. & Knowles, D. P. ( 1998; ). Monoclonal antibody F89/160.1.5 defines a conserved epitope on the ruminant prion protein. J Clin Microbiol 36, 1750–1755.
    [Google Scholar]
  22. Parchi, P., Castellani, R., Capellari, S. & 9 other authors ( 1996; ). Molecular basis of phenotypic variability in sporadic Creutzfeldt-Jakob disease. Ann Neurol 39, 767–778.[CrossRef]
    [Google Scholar]
  23. Parchi, P., Capellari, S., Chen, S. G. & 8 other authors ( 1997; ). Typing prion isoforms. Nature 386, 232–234.[CrossRef]
    [Google Scholar]
  24. Parchi, P., Chen, S. G., Brown, P. & 9 other authors ( 1998; ). Different patterns of truncated prion protein fragments correlate with distinct phenotypes in P102L Gerstmann-Sträussler-Scheinker disease. Proc Natl Acad Sci U S A 95, 8322–8327.[CrossRef]
    [Google Scholar]
  25. Parchi, P., Zou, W., Wang, W. & 10 other authors ( 2000; ). Genetic influence on the structural variations of the abnormal prion protein. Proc Natl Acad Sci U S A 97, 10168–10172.[CrossRef]
    [Google Scholar]
  26. Piccardo, P., Ghetti, B., Dickson, D. W. & other authors ( 1995; ). Gerstmann-Sträussler-Scheinker disease (PRNP P102L): amyloid deposits are best recognized by antibodies directed to epitopes in PrP region 90-165. J Neuropathol Exp Neurol 54, 790–801.[CrossRef]
    [Google Scholar]
  27. Piccardo, P., Seiler, C., Dlouhy, S. R. & 7 other authors ( 1996; ). Proteinase-K-resistant prion protein isoforms in Gerstmann-Sträussler-Scheinker disease (Indiana kindred). J Neuropathol Exp Neurol 55, 1157–1163.[CrossRef]
    [Google Scholar]
  28. Piccardo, P., Liepnieks, J. J., William, A. & 12 other authors ( 2001; ). Prion proteins with different conformations accumulate in Gerstmann-Sträussler-Scheinker disease caused by A117V and F198S mutations. Am J Pathol 158, 2201–2207.[CrossRef]
    [Google Scholar]
  29. Prusiner, S. B. ( 1998; ). Prions. Proc Natl Acad Sci U S A 95, 13363–13383.[CrossRef]
    [Google Scholar]
  30. Salmona, M., Morbin, M., Massignan, T. & 14 other authors ( 2003; ). Structural properties of Gerstmann-Sträussler-Scheinker disease amyloid protein. J Biol Chem 278, 48146–48153.[CrossRef]
    [Google Scholar]
  31. Sigurdson, C. J. & Miller, M. W. ( 2003; ). Other animal prion diseases. Br Med Bull 66, 199–212.[CrossRef]
    [Google Scholar]
  32. Somerville, R. A. & Ritchie, L. A. ( 1990; ). Differential glycosylation of the protein (PrP) forming scrapie-associated fibrils. J Gen Virol 71, 833–839.[CrossRef]
    [Google Scholar]
  33. Stahl, N., Baldwin, M. A., Teplow, D. B., Hood, L., Gibson, B. W., Burlingame, A. L. & Prusiner, S. B. ( 1993; ). Structural studies of the scrapie prion protein using mass spectrometry and amino acid sequencing. Biochemistry 32, 1991–2002.[CrossRef]
    [Google Scholar]
  34. Stockman, S. ( 1913; ). Scrapie: an obscure disease of sheep. J Comp Pathol 26, 317–327.[CrossRef]
    [Google Scholar]
  35. Tagliavini, F., Prelli, F., Ghiso, J., Bugiani, O., Serban, D., Prusiner, S. B., Farlow, M. R., Ghetti, B. & Frangione, B. ( 1991; ). 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 10, 513–519.
    [Google Scholar]
  36. Tagliavini, F., Prelli, F., Porro, M. & 7 other authors ( 1994; ). Amyloid fibrils in Gerstmann-Sträussler-Scheinker disease (Indiana and Swedish kindreds) express only PrP peptides encoded by the mutant allele. Cell 79, 695–703.[CrossRef]
    [Google Scholar]
  37. Tagliavini, F., Lievens, P. M.-J., Tranchant, C. & 12 other authors ( 2001; ). A 7-kDa prion protein (PrP) fragment, an integral component of the PrP region required for infectivity, is the major amyloid protein in Gerstmann-Sträussler-Scheinker disease A117V. J Biol Chem 276, 6009–6015.[CrossRef]
    [Google Scholar]
  38. Thuring, C. M. A., Erkens, J. H. F., Jacobs, J. G. & 8 other authors ( 2004; ). Discrimination between scrapie and bovine spongiform encephalopathy in sheep by molecular size, immunoreactivity, and glycoprofile of prion protein. J Clin Microbiol 42, 972–980.[CrossRef]
    [Google Scholar]
  39. Van Everbroeck, B., O'Rourke, K. I. & Cras, P. ( 1999; ). Immunoreactivity of the monoclonal antibody F89/160.1.5 for the human prion protein. Eur J Histochem 43, 335–338.
    [Google Scholar]
  40. van Keulen, L. J., Schreuder, B. E., Meloen, R. H., Poelen-van den Berg, M., Mooij-Harkes, G., Vromans, M. E. & Langeveld, J. P. ( 1995; ). Immunohistochemical detection and localization of prion protein in brain tissue of sheep with natural scrapie. Vet Pathol 32, 299–308.[CrossRef]
    [Google Scholar]
  41. Vorberg, I., Buschmann, A., Harmeyer, S., Saalmüller, A., Pfaff, E. & Groschup, M. H. ( 1999; ). A novel epitope for the specific detection of exogenous prion proteins in transgenic mice and transfected murine cell lines. Virology 255, 26–31.[CrossRef]
    [Google Scholar]
  42. Yadavalli, R., Guttmann, R. P., Seward, T., Centers, A. P., Williamson, R. A. & Telling, G. C. ( 2004; ). Calpain-dependent endoproteolytic cleavage of PrPSc modulates scrapie prion propagation. J Biol Chem 279, 21948–21956.[CrossRef]
    [Google Scholar]
  43. Young, K., Piccardo, P., Dlouhy, S. R., Bugiani, O., Tagliavini, F. & Ghetti, B. ( 1999; ). The human genetic prion diseases. In Prions: Molecular and Cellular Biology, pp. 139–175. Edited by D. A. Harris. Wymondham, UK: Horizon Scientific Press.
  44. Yuan, J., Kinter, M., McGeehan, J., Perry, G., Kneale, G., Gambetti, P. & Zou, W. Q. ( 2005; ). Concealment of epitope by reduction and alkylation in prion protein. Biochem Biophys Res Commun 326, 652–659.[CrossRef]
    [Google Scholar]
  45. Zhao, H., Klingeborn, M., Simonsson, M. & Linné, T. ( 2006; ). Proteolytic cleavage and shedding of the bovine prion protein in two cell culture systems. Virus Res 115, 43–55.[CrossRef]
    [Google Scholar]
  46. Zou, W.-Q., Capellari, S., Parchi, P., Sy, M. S., Gambetti, P. & Chen, S.-G. ( 2003; ). Identification of novel proteinase K-resistant C-terminal fragments of PrP in Creutzfeldt-Jakob disease. J Biol Chem 278, 40429–40436.[CrossRef]
    [Google Scholar]
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