Summary
A central event in the transmission and pathogenesis of transmissible spongiform encephalopathy diseases is the misfolding of the prion protein. Considerable progress has been made in our understanding of this misfolding event through the development of cell-free assays that mimic the molecular features of prion propagation. This chapter reviews the contribution of cell-free assays to our understanding of prion propagation.
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References
Kocisko, D. A., Come, J. H., Priola, S. A., Chesebro, B., Raymond, G. J., Lansbury, P. T., and Caughey, B. (1994) Cell-free formation of protease-resistant prion protein. Nature 370, 471–4.
Caughey, B., Raymond, G. J., Priola, S. A., Kocisko, D. A., Race, R. E., Bessen, R. A., Lansbury, P. T., Jr., and Chesebro, B. (1999) Methods for studying prion protein (PrP) metabolism and the formation of protease-resistant PrP in cell culture and cell-free systems. An update. Mol Biotechnol 13, 45–55.
Hill, A. F., Antoniou, M., and Collinge, J. (1999) Protease-resistant prion protein produced in vitro lacks detectable infectivity. J Gen Virol 80 (1), 11–4.
Kocisko, D. A., Priola, S. A., Raymond, G. J., Chesebro, B., Lansbury, P. T., Jr., and Caughey, B. (1995) Species specificity in the cell-free conversion of prion protein to protease-resistant forms: a model for the scrapie species barrier. Proc Natl Acad Sci U S A 92, 3923–7.
Bessen, R. A., Kocisko, D. A., Raymond, G. J., Nandan, S., Lansbury, P. T., and Caughey, B. (1995) Non-genetic propagation of strain-specific properties of scrapie prion protein. Nature 375, 698–700.
Raymond, G. J., Bossers, A., Raymond, L. D., O'Rourke, K. I., McHolland, L. E., Bryant, P. K., 3rd, Miller, M. W., Williams, E. S., Smits, M., and Caughey, B. (2000) Evidence of a molecular barrier limiting susceptibility of humans, cattle and sheep to chronic wasting disease. EMBO J 19, 4425–30.
Raymond, G. J., Hope, J., Kocisko, D. A., Priola, S. A., Raymond, L. D., Bossers, A., Ironside, J., Will, R. G., Chen, S. G., Petersen, R. B., Gambetti, P., Rubenstein, R., Smits, M. A., Lansbury, P. T., Jr., and Caughey, B. (1997) Molecular assessment of the potential transmissi-bilities of BSE and scrapie to humans. Nature 388, 285–8.
Priola, S. A., Chabry, J., and Chan, K. (2001) Efficient conversion of normal prion protein (PrP) by abnormal hamster PrP is determined by homology at amino acid residue 155. J Virol 75, 4673–80.
Priola, S. A., and Lawson, V. A. (2001) Glycosylation influences cross-species formation of protease-resistant prion protein. EMBO J 20, 6692–9.
Vorberg, I., and Priola, S. A. (2002) Molecular basis of scrapie strain glycoform variation. J Biol Chem 277, 36775–81.
Horiuchi, M., and Caughey, B. (1999) Specific binding of normal prion protein to the scrapie form via a localized domain initiates its conversion to the protease-resistant state. EMBO J 18, 3193–203.
Caughey, B., Raymond, G. J., Ernst, D., and Race, R. E. (1991) N-terminal truncation of the scrapie-associated form of PrP by lysosomal protease(s): implications regarding the site of conversion of PrP to the protease-resistant state. J Virol 65, 6597–603.
Baron, G. S., Wehrly, K., Dorward, D. W., Chesebro, B., and Caughey, B. (2002) Conversion of raft associated prion protein to the protease-resistant state requires insertion of PrP-res (PrP(Sc) ) into contiguous membranes. EMBO J 21, 1031–40.
Caughey, W. S., Raymond, L. D., Horiuchi, M., and Caughey, B. (1998) Inhibition of protease-resistant prion protein formation by porphyrins and phthalocyanines. Proc Natl Acad Sci U S A 95, 12117–22.
Maxson, L., Wong, C., Herrmann, L. M., Caughey, B., and Baron, G. S. (2003) A solid-phase assay for identification of modulators of prion protein interactions. Anal Biochem 323, 54–64.
Kirby, L., Birkett, C. R., Rudyk, H., Gilbert, I. H., and Hope, J. (2003) In vitro cell-free conversion of bacterial recombinant PrP to PrPres as a model for conversion. J Gen Virol 84, 1013–20.
Silveira, J. R., Raymond, G. J., Hughson, A. G., Race, R. E., Sim, V. L., Hayes, S. F., and Caughey, B. (2005) The most infectious prion protein particles. Nature 437, 257–61.
Saborio, G. P., Permanne, B., and Soto, C. (2001) Sensitive detection of pathological prion protein by cyclic amplification of protein misfolding. Nature 411, 810–3.
Castilla, J., Saa, P., Morales, R., Abid, K., Maundrell, K., and Soto, C. (2006) Protein misfold-ing cyclic amplification for diagnosis and prion propagation studies. Methods Enzymol 412, 3–21.
Castilla, J., Saa, P., Hetz, C., and Soto, C. (2005) In vitro generation of infectious scrapie pri-ons. Cell 121, 195–206.
Bieschke, J., Weber, P., Sarafoff, N., Beekes, M., Giese, A., and Kretzschmar, H. (2004) Autocatalytic self-propagation of misfolded prion protein. Proc Natl Acad Sci U S A 101, 12207–11.
Castilla, J., Saa, P., and Soto, C. (2005) Detection of prions in blood. Nat Med 11, 982–5.
Wong, C., Xiong, L. W., Horiuchi, M., Raymond, L., Wehrly, K., Chesebro, B., and Caughey, B. (2001) Sulfated glycans and elevated temperature stimulate PrP(Sc)-dependent cell-free formation of protease-resistant prion protein. EMBO J 20, 377–86.
Deleault, N. R., Geoghegan, J. C., Nishina, K., Kascsak, R., Williamson, R. A., and Supattapone, S. (2005) Protease-resistant prion protein amplification reconstituted with partially purified substrates and synthetic polyanions. J Biol Chem 280, 26873–9.
Supattapone, S. (2004) Prion protein conversion in vitro. J Mol Med 82, 348–56.
Legname, G., Baskakov, I. V. , Nguyen, H. O., Riesner, D., Cohen, F. E., DeArmond, S. J., and Prusiner, S. B. (2004) Synthetic mammalian prions. Science 305, 673–6.
Nazor, K. E., Kuhn, F., Seward, T., Green, M., Zwald, D., Purro, M., Schmid, J., Biffiger, K., Power, A. M., Oesch, B., Raeber, A. J., and Telling, G. C. (2005) Immunodetection of disease-associated mutant PrP, which accelerates disease in GSS transgenic mice. EMBO J 24, 2472–80.
Zou, W. Q., Capellari, S., Parchi, P., Sy, M. S., Gambetti, P., and Chen, S. G. (2003) Identification of novel proteinase K-resistant C-terminal fragments of PrP in Creutzfeldt-Jakob disease. J Biol Chem 278, 40429–36.
Lawson, V. A., Priola, S. A., Meade-White, K., Lawson, M., and Chesebro, B. (2004) Flexible N-terminal region of prion protein influences conformation of protease-resistant prion protein isoforms associated with cross-species scrapie infection in vivo and in vitro. J Biol Chem 279, 13689–95.
Bocharova, O. V. , Breydo, L., Salnikov, V. V., Gill, A. C., and Baskakov, I. V. (2005) Synthetic prions generated in vitro are similar to a newly identified subpopulation of PrPSc from sporadic Creutzfeldt-Jakob disease. Protein Sci 14, 1222–32.
Moore, R. A., Herzog, C., Errett, J., Kocisko, D. A., Arnold, K. M., Hayes, S. F., and Priola, S. A. (2006) Octapeptide repeat insertions increase the rate of protease-resistant prion protein formation. Protein Sci 15, 609–19.
Baskakov, I., Disterer, P., Breydo, L., Shaw, M., Gill, A., James, W., and Tahiri-Alaoui, A. (2005) The presence of valine at residue 129 in human prion protein accelerates amyloid formation. FEBS Lett 579, 2589–96.
Acknowledgments
This work was supported by the National Health and Medical Research Council (NHMRC) of Australia fellowship 209163 and project grants 400229 and 454546.
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© 2008 Humana Press, a part of Springer Science + Business Media, LLC
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Lawson, V.A. (2008). Understanding the Nature of Prion Diseases Using Cell-free Assays. In: Hill, A.F. (eds) Prion Protein Protocols. Methods in Molecular Biology™, vol 459. Humana Press. https://doi.org/10.1007/978-1-59745-234-2_7
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DOI: https://doi.org/10.1007/978-1-59745-234-2_7
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