Prion Proteins pp 121-133 | Cite as

Prion Seeded Conversion and Amplification Assays

  • Christina D. Orrú
  • Byron CaugheyEmail author
Part of the Topics in Current Chemistry book series (TOPCURRCHEM, volume 305)


The conversion of the normal prion protein (PrPC) into its misfolded, aggregation-prone and infectious (prion) isoform is central to the progression of transmissible spongiform encephalopathies (TSEs) or prion diseases. Since the initial development of a cell free PrP conversion reaction, striking progress has been made in the development of much more continuous prion-induced conversion and amplification reactions. These studies have provided major insights into the molecular underpinnings of prion propagation and enabled the development of ultra-sensitive tests for prions and prion disease diagnosis. This chapter will provide an overview of such reactions and the practical and fundamental consequences of their development.


Prion composition PMCA QuIC ASA Transmissible Spongiform Ecephalopathies 



We thank Dr. Jason M. Wilham for designing the figures and Anita Mora for graphics assistance.


  1. 1.
    Caughey B, Baron GS, Chesebro B, Jeffrey M (2009) Annu Rev Biochem 78:177CrossRefGoogle Scholar
  2. 2.
    Kocisko DA, Come JH, Priola SA, Chesebro B, Raymond GJ, Lansbury PT, Caughey B (1994) Nature 370:471CrossRefGoogle Scholar
  3. 3.
    Bossers A, Belt PBGM, Raymond GJ, Caughey B, de Vries R, Smits MA (1997) Proc Natl Acad Sci USA 94:4931CrossRefGoogle Scholar
  4. 4.
    Horiuchi M, Priola SA, Chabry J, Caughey B (2000) Proc Natl Acad Sci USA 97:5836CrossRefGoogle Scholar
  5. 5.
    Kocisko DA, Priola SA, Raymond GJ, Chesebro B, Lansbury PT Jr, Caughey B (1995) Proc Natl Acad Sci USA 92:3923CrossRefGoogle Scholar
  6. 6.
    Raymond GJ, Bossers A, Raymond LD, O’Rourke KI, McHolland LE, Bryant PK III, Miller MW, Williams ES, Smits M, Caughey B (2000) EMBO J 19:4425CrossRefGoogle Scholar
  7. 7.
    Raymond GJ, Hope J, Kocisko DA, Priola SA, Raymond LD, Bossers A, Ironside J, Will RG, Chen SG, Petersen RB, Gambetti P, Rubenstein R, Smits MA, Lansbury PT Jr, Caughey B (1997) Nature 388:285CrossRefGoogle Scholar
  8. 8.
    Bessen RA, Kocisko DA, Raymond GJ, Nandan S, Lansbury PT Jr, Caughey B (1995) Nature 375:698CrossRefGoogle Scholar
  9. 9.
    Hill AF, Antoniou M, Collinge J (1999) J Gen Virol 80:11Google Scholar
  10. 10.
    Caughey B (2003) Br Med Bull 66:109CrossRefGoogle Scholar
  11. 11.
    Saborio GP, Permanne B, Soto C (2001) Nature 411:810CrossRefGoogle Scholar
  12. 12.
    Soto C, Anderes L, Suardi S, Cardone F, Castilla J, Frossard MJ, Peano S, Saa P, Limido L, Carbonatto M, Ironside J, Torres JM, Pocchiari M, Tagliavini F (2005) FEBS Lett 579:638CrossRefGoogle Scholar
  13. 13.
    Thorne L, Terry LA (2008) J Gen Virol 89:3177CrossRefGoogle Scholar
  14. 14.
    Kurt TD, Perrott MR, Wilusz CJ, Wilusz J, Supattapone S, Telling GC, Zabel MD, Hoover EA (2007) J Virol 81:9605CrossRefGoogle Scholar
  15. 15.
    Murayama Y, Yoshioka M, Okada H, Takata M, Yokoyama T, Mohri S (2007) J Gen Virol 88:2890CrossRefGoogle Scholar
  16. 16.
    Jones M, Peden AH, Prowse CV, Groner A, Manson JC, Turner ML, Ironside JW, Macgregor IR, Head MW (2007) J Pathol 213:21CrossRefGoogle Scholar
  17. 17.
    Jones M, Peden AH, Yull H, Wight D, Bishop MT, Prowse CV, Turner ML, Ironside JW, Macgregor IR, Head MW (2009) Transfusion 49:376CrossRefGoogle Scholar
  18. 18.
    Pastrana MA, Sajnani G, Onisko B, Castilla J, Morales R, Soto C, Requena JR (2006) Biochemistry 45:15710CrossRefGoogle Scholar
  19. 19.
    Castilla J, Saa P, Morales R, Abid K, Maundrell K, Soto C (2006) Meth Enzymol 412:3CrossRefGoogle Scholar
  20. 20.
    Fujihara A, Atarashi R, Fuse T, Ubagai K, Nakagaki T, Yamaguchi N, Ishibashi D, Katamine S, Nishida N (2009) FEBS J 276:2841CrossRefGoogle Scholar
  21. 21.
    Saa P, Castilla J, Soto C (2006) Science 313:92CrossRefGoogle Scholar
  22. 22.
    Tattum MH, Jones S, Pal S, Collinge J, Jackson GS (2010) Transfusion 50:996CrossRefGoogle Scholar
  23. 23.
    Nichols TA, Pulford B, Wyckoff AC, Meyerett C, Michel B, Gertig K, Hoover EA, Jewell JE, Telling GC, Zabel MD (2009) Prion 3:171CrossRefGoogle Scholar
  24. 24.
    Saa P, Castilla J, Soto C (2006) J Biol Chem 281:35245CrossRefGoogle Scholar
  25. 25.
    Chen B, Morales R, Barria MA, Soto C (2010) Nat Methods 7:519CrossRefGoogle Scholar
  26. 26.
    Atarashi R, Moore RA, Sim VL, Hughson AG, Dorward DW, Onwubiko HA, Priola SA, Caughey B (2007) Nat Methods 4:645CrossRefGoogle Scholar
  27. 27.
    Legname G, Baskakov IV, Nguyen HO, Riesner D, Cohen FE, DeArmond SJ, Prusiner SB (2004) Science 305:673CrossRefGoogle Scholar
  28. 28.
    Colby DW, Wain R, Baskakov IV, Legname G, Palmer CG, Nguyen HO, Lemus A, Cohen FE, DeArmond SJ, Prusiner SB (2010) PLoS Pathog 6:e1000736CrossRefGoogle Scholar
  29. 29.
    Castilla J, Saa P, Hetz C, Soto C (2005) Cell 121:195CrossRefGoogle Scholar
  30. 30.
    Weber P, Giese A, Piening N, Mitteregger G, Thomzig A, Beekes M, Kretzschmar HA (2006) Proc Natl Acad Sci USA 103:15818CrossRefGoogle Scholar
  31. 31.
    Weber P, Giese A, Piening N, Mitteregger G, Thomzig A, Beekes M, Kretzschmar HA (2007) Vet Microbiol 123:346CrossRefGoogle Scholar
  32. 32.
    Weber P, Reznicek L, Mitteregger G, Kretzschmar H, Giese A (2008) Biochem Biophys Res Commun 369:924CrossRefGoogle Scholar
  33. 33.
    Green KM, Castilla J, Seward TS, Napier DL, Jewell JE, Soto C, Telling GC (2008) PLoS Pathog 4:e1000139CrossRefGoogle Scholar
  34. 34.
    Deleault NR, Harris BT, Rees JR, Supattapone S (2007) Proc Natl Acad Sci USA 104:9741CrossRefGoogle Scholar
  35. 35.
    Deleault NR, Geoghegan JC, Nishina K, Kascsak R, Williamson RA, Supattapone S (2005) J Biol Chem 280:26873CrossRefGoogle Scholar
  36. 36.
    Geoghegan JC, Valdes PA, Orem NR, Deleault NR, Williamson RA, Harris BT, Supattapone S (2007) J Biol Chem 282:36341CrossRefGoogle Scholar
  37. 37.
    Wang F, Wang X, Yuan CG, Ma J (2010) Science 327:1132CrossRefGoogle Scholar
  38. 38.
    Kim JI, Cali I, Surewicz K, Kong Q, Raymond GJ, Atarashi R, Race B, Qing L, Gambetti P, Caughey B, Surewicz WK (2010) J Biol Chem 285:14083CrossRefGoogle Scholar
  39. 39.
    Abid K, Morales R, Soto C (2010) FEBS Lett 584:2409CrossRefGoogle Scholar
  40. 40.
    Deleault NR, Kascsak R, Geoghegan JC, Supattapone S (2010) Biochemistry 49:3928CrossRefGoogle Scholar
  41. 41.
    Nishina KA, Deleault NR, Mahal SP, Baskakov I, Luhrs T, Riek R, Supattapone S (2006) Biochemistry 45:14129CrossRefGoogle Scholar
  42. 42.
    Piro JR, Harris BT, Nishina K, Soto C, Morales R, Rees JR, Supattapone S (2009) J Virol 83:5321CrossRefGoogle Scholar
  43. 43.
    Bessen RA, Marsh RF (1994) J Virol 68:7859Google Scholar
  44. 44.
    Collinge J, Clarke AR (2007) Science 318:930CrossRefGoogle Scholar
  45. 45.
    Morales R, Abid K, Soto C (2007) Biochim Biophys Acta 1772:681Google Scholar
  46. 46.
    Telling GC, Parchi P, DeArmond SJ, Cortelli P, Montagna P, Gabizon R, Mastrianni J, Lugaresi E, Gambetti P, Prusiner SB (1996) Science 274:2079CrossRefGoogle Scholar
  47. 47.
    Castilla J, Morales R, Saa P, Barria M, Gambetti P, Soto C (2008) EMBO J 27:2557CrossRefGoogle Scholar
  48. 48.
    Castilla J, Gonzalez-Romero D, Saa P, Morales R, De CJ, Soto C (2008) Cell 134:757CrossRefGoogle Scholar
  49. 49.
    Barria MA, Mukherjee A, Gonzalez-Romero D, Morales R, Soto C (2009) PLoS Pathog 5:e1000421CrossRefGoogle Scholar
  50. 50.
    Colby DW, Zhang Q, Wang S, Groth D, Legname G, Riesner D, Prusiner SB (2007) Proc Natl Acad Sci USA 104:20914CrossRefGoogle Scholar
  51. 51.
    LeVine H III (1999) Meth Enzymol 309:274CrossRefGoogle Scholar
  52. 52.
    Atarashi R, Satoh K, Sano K, Fuse T, Yamaguchi N, Ishibashi D, Matsubara T, Nakagaki T, Yamanaka H, Shirabe S, Yamada M, Mizusawa H, Kitamoto T, Klug G, McGlade A, Collins SJ, Nishida N (2011) Nat Med 17:175CrossRefGoogle Scholar
  53. 53.
    Atarashi R, Wilham JM, Christensen L, Hughson AG, Moore RA, Johnson LM, Onwubiko HA, Priola SA, Caughey B (2008) Nat Methods 5:211CrossRefGoogle Scholar
  54. 54.
    Orrú CD, Wilham JM, Hughson AG, Raymond LD, McNally KL, Bossers A, Ligios C, Caughey B (2009) Protein Eng Des Sel 22:515CrossRefGoogle Scholar
  55. 55.
    Wilham JM, Orrú CD, Bessen RA, Atarashi R, Sano K, Race B, Meade-White KD, Taubner LM, Timmes A, Caughey B (2010) PLoS Pathog 6:e1001217CrossRefGoogle Scholar
  56. 56.
    Bessen RA, Shearin H, Martinka S, Boharski R, Lowe D, Wilham JM, Caughey B, Wiley JA (2010) PLoS Pathog 6:e1000837CrossRefGoogle Scholar
  57. 57.
    Dougherty RM (1964) Animal virus titration techniques. In: Harris RJC (ed) Techniques in experimental virology. Academic, New York, p 183Google Scholar
  58. 58.
    Reed LJ, Muench H (1938) Am J Hyg 27:493Google Scholar
  59. 59.
    Gregori L, McCombie N, Palmer D, Birch P, Sowemimo-Coker SO, Giulivi A, Rohwer RG (2004) Lancet 364:529CrossRefGoogle Scholar
  60. 60.
    Trieschmann L, Navarrete SA, Kaschig K, Torkler S, Maas E, Schatzl H, Bohm G (2005) BMC Biotechnol 5:26CrossRefGoogle Scholar
  61. 61.
    Orrú CD, Wilham JM, Raymond LD, Kuhn F, Schroeder B, Raebet A, Coughey B (2011) mBio 2(3):c00078–11Google Scholar
  62. 62.
    Edgeworth JA, Farmer M, Sicilia A, Tavares P, Beck J, Campbell T, Lowe J, Mead S, Rudge P, Collinge J, Jackson GS (2011) Lancet 377:487CrossRefGoogle Scholar
  63. 63.
    Miller MB, Supattapone S (2011) J Virol 85:2813CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin-Heidelberg 2011

Authors and Affiliations

  1. 1.Laboratory of Persistent Viral Diseases, Rocky Mountain LaboratoriesNational Institute of Allergy and Infectious Diseases, National Institutes of HealthHamiltonUSA

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