Abstract
In the past two decades, thoroughly standardized mouse incubation time and brain lesion profile scoring assays have been developed to discriminate between prion strains. However, in these mouse infection experiments, large numbers of animals (about 20 mice/line) from three different highly inbred mouse lines (C57Bl, VM95, RIII), plus their intercrosses, need to be infected, and their brain tissues subsequently examined (1-8). Although results obtained are highly reliable, the effort and time needed for conducting these experiments are considerable. Therefore, alternative criteria and techniques have been developed to characterize transmissible spongiform encephalopathy (TSE) agents. Prion infections are accompanied by the accumulation of an abnormal isoform (designated PrPSc) of normal host-encoded prion protein (PrPC). Both isoforms have the same amino acid sequence and molecular mass, but differ significantly in their three-dimensional structure and biochemical characteristics. The three-dimensional structure of PrPC is characterized by a high ?-helical content (9); all or part of it undergoes a posttranslational modification to β-sheet in PrPSc (10-12). Although PrPC (33-35 kDa) is completely hydrolyzed by protease treatment, PrPSc is partially resistant to proteinase K(PK) as 62 N-terminal amino acids are cleaved, leaving a core fragment of approx 141 amino acids (27-30 kDa) unhydrolyzed (13).
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References
Fraser, H. and Dickinson, A. G. (1973) Scrapie in mice. Agent-strain differences in the distribution and intensity of grey matter vacuolation. J. Comp. Pathol. 83, 29–40.
Outram, G. W., Fraser, H., and Wilson, D. T. (1973) Scrapie in mice. Some effects on the brain lesion profile of ME7 agent due to genotype of donor, route of injection and genotype of recipient. J. Comp.Pathol. 83, 19–28.
Fraser, H. (1976) The pathology of a natural and experimental scrapie. Frontiers Biol. 44, 267–305.
Bruce, M. E., McBride, P. A., Jeffrey, M., and Scott, J. R. (1994) PrP in pathology and pathogenesis in scrapie-infected mice. Mol. Neurobiol. 8, 105–112.
Fraser, H., Bruce, M. E., and McConnell, I. (1991) Murine scrapie strains, BSE models and genetics, in Sub-Acute Spongiform Encephalopathies (Bradley, R., Savey, M. and Marchant, B. eds.), Kluwer, Dordrecht, The Netherlands, pp. 131–136.
Bruce, M., Chree, A., McConnell, I., Brown, K., and Fraser, H. (1996) Transmission and strain typing studies of scrapie and bovine spongiform encephalopathy, in Transmissible Subacute Spongiform Encephalopathies: Prion Diseases (Court, L. and Dodet, B., eds.), Elsevier, Paris. pp. 259–262.
Bruce, M. E. and Dickinson, A. G. (1987) Biological evidence that scrapie agent has an independent genome. J. Gen.Virol 68, 79–89.
Bruce, M. E. (1996) Strain typing studies of scrapie and BSE, in Prion Diseases (Baker, H. F. and Ridley, R. M., eds.), Humana, Totowa, NJ pp. 223–236.
Riek, R., Hornemann, S., Wider, G., Billeter, M., Glockshuber, R., and Wuthrich, K. (1996) NMR structure of the mouse prion protein domain PrP(121-321). Nature 382, 180–182.
Pan, K. M., Baldwin, M., Nguyen, J., Gasset, M., Serban, A., Groth, D., et al. (1993) Conversion of alpha-helices into beta-sheets features in the formation of the scrapie prion proteins. Proc. Natl. Acad. Sci. USA 90, 10,962–10,966.
Pergami, P., Jaffe, H., and Safar J. (1996) Semipreparative chromatographic method to purify the normal cellular isoform of the prion protein in nondenatured form. Analyt. Biochem. 236, 63–73.
Safar, J., Roller, P. P., Gajdusek, D. C., and Gibbs, C. J. Jr., (1993) Conformational transitions, dissociation, and unfolding of scrapie amyloid (prion) protein. J. Biol. Chem. 268, 20,276–20,284.
Oesch, B., Westaway, D., Walchli, M., McKinley, M. P., Kent, S. B., Aebersold, R. et al. (1985) A cellular gene encodes scrapie PrP 27-30 protein. Cell 40; 735–746.
Kascsak, R. J., Rubenstein, R., Merz, P. A., Carp, R. I., Wisniewski, H. M., and Diringer, H. (1985) Biochemical differences among scrapie-associated fibrils support the biological diversity of scrapie agents. J. Gen. Virol. 66, 1715–1722.
Kascsak, R. J., Rubenstein, R., Merz, P. A., Carp, R. I., Robakis, N. K., Wisniewski, H. M., and Diringer, H. (1986) Immunological comparison of scrapie-associated fibrils isolated from animals infected with four different scrapie strains. J. Virol. 59, 676–683.
Bessen, R. A. and Marsh, R. F. (1992) Biochemical and physical properties of the prion protein from two strains of the transmissible mink encephalopathy agent. J. Virol. 66, 2096–2101.
Marsh, R. F. and Bessen, R. A. (1994) Physicochemical and biological characterizations of distinct strains of the transmissible mink encephalopathy agent. Phil. Trans. Roy. Soc. London-Series B: Biol. Sci. 343, 413–414.
Collinge, J., Sidle, K. C., Meads, J., Ironside, J., and Hill, A. F. (1996) Molecular analysis of prion strain variation and the aetiology of ‘new variant’ CJD. Nature 383, 685–690.
Parchi, P., Capellari, S., Chen, S. G., Petersen, R. B., Gambetti, P., Kopp, N. et al. (1997) Typing prion isoforms. Nature 386, 232–234.
Cardone, F., Liu, Q. G., Petraroli, R., Ladogana, A., D’Alessandro, M., Arpino, C., et al. (1999) Prion protein glycotype analysis in familial and sporadic Creutzfeldt-Jakob disease patients. Brain Res. Bull. 49, 429–433.
Wadsworth, J. D., Hill, A. F., Joiner, S., Jackson, G. S., Clarke, A. R., and Collinge, J. (1999) Strain-specific prion-protein conformation determined by metal ions. Nat. Cell Biol. 1, 55–59.
Endo, T., Groth, D., Prusiner, S. B., and Kobata, A. (1989) Diversity of oligosaccharide structures linked to asparagines of the scrapie prion protein. Biochemistry 28, 8380–8388.
Somerville, R. A., Chong, A., Mulqueen, O. U., Birkett, C. R., Wood, S. C., and Hope, J. (1997) Biochemical typing of scrapie strains. Nature 386, 564–564.
Hope, J., Reekie, L. J., Hunter, N., Multhaup, G., Beyreuther, K., White, H., et al. (1988) Fibrils from brains of cows with new cattle disease contain scrapie-associated protein. Nature 336, 390–392.
Somerville, R. A. (1999) Host and transmissible spongiform encephalopathy agent strain control glycosylation of PrP. J. Gen. Virol. 80, 1865–1872.
Kuczius, T., Haist, I., and Groschup, M. H. (1998) Molecular analysis of bovine spongiform encephalopathy and scrapie strain variation. J. Infect. Dis. 178, 693–699.
Baron, T. G., Madec, J. Y., and Calavas, D. (1999) Similar signature of the prion protein in natural sheep scrapie and bovine spongiform encephalopathy-linked diseases. J. Clin. Microbiol. 37, 3701–3704.
Sweeney, T., Kuczius, T., McElroy, M., Gomerez-Parada, M., Groschup, M. H. (2000) Molecular Analysis of Irish scrapie cases. J. Gen. Virol. 6, 1621–1627.
Hill, A. F., Butterworth, R. J., Joiner, S., Jackson, G., Rossor, M. N., Thomas, D. J, et al. (1999) Investigation of variant Creutzfeldt-Jakob disease and other human prion diseases with tonsil biopsy samples. Lancet 353, 183–189.
Buschmann, A., Kuczius, T., Bodemer, W., and Groschup, M. H. (1998) Cellular prion proteins of mammalian species display an intrinsic partial proteinase K resistance. Biochem. Biophys. Res. Commun. 253, 693–702.
Groschup, M. H. and E. Pfaff (1993) Studies on a species-specific epitope in murine, ovine and bovine prion protein. J. Gen. Virol 74, 1451–1456.
Kascsak, R. J., Rubenstein, R., Merz, P. A., Tonna-DeMasi, M., Fersko, R., Carp, R. I., et al. (1987) Mouse polyclonal and monoclonal antibody to scrapie-associated fibril proteins. J. Virol. 61, 3688–3693.
Vorberg, I., Buschmann, A., Harmeyer, S., Saalmuller, A., Pfaff, E., and 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.
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Groschup, M.H., Junghans, F., Eiden, M., Kuczius, T. (2001). Characterization of Bovine Spongiform Encephalopathy and Scrapie Strains/Isolates by Immunochemical Analysis of PrPSc. In: Baker, H.F. (eds) Molecular Pathology of the Prions. Methods in Molecular Medicine™, vol 59. Humana Press. https://doi.org/10.1385/1-59259-134-5:71
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DOI: https://doi.org/10.1385/1-59259-134-5:71
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