Design of a Coiled-Coil-based Model Peptide System to Explore the Fundamentals of Amyloid Fibril Formation

  • Michel O. Steinmetz
  • Carlos García-Echeverría
  • Richard A. Kammerer


Protein deposition as amyloid fibrils underlies more than twenty severely debilitating human disorders. Interestingly, recent studies suggest that all peptides and proteins possess an intrinsic ability to assemble into amyloid fibrils similar to those observed in disease states. The common properties and characteristics of amyloid aggregates thus offer the prospect that simple model systems can be used to systematically assess the factors that predispose a native protein to form amyloid fibrils and understand the origin and progression of fatal disorders associated with amyloid formation. Here, we report the de novo design of a 17-residue peptide model system, referred to as ccβ, which forms a protein-like coiled-coil structure under ambient solution conditions but can be easily converted into amyloid fibrils by raising the temperature. Oxidation of methionine residues at selected hydrophobic positions completely abolished amyloid fibril formation of the peptide while not interfering with its coiled-coil structure. This finding indicates that a small number of site-specific hydrophobic interactions can play a major role in the packing of polypeptide chain segments within amyloid fibrils. The simplicity and characteristics of the ccβ system make it highly suitable for probing molecular details of the assembly of amyloid structures.

Key words

Coiled coil de novo design amyloid 



circular dichroism






Grand average hydrophaty


high-performance liquid chromatography


matrix-assisted laser-desorption ionization time-of-flight mass spectroscopy




tris(alkoxy)benzylamide linker


phosphate buffered saline


polyethylene glycol


transmission electron microscope


O-(1,2-dihydro-2-oxo-pyridyl)-N, N, N′,N′-tetramethyluronium tetrafluoroborate


trifluoroacetic acid




Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Aguzzi, A., Haass, C. 2003Science302814Google Scholar
  2. Albericio, F., Kneib-Cordonier, N., Biancalana, S., Gera, L., Masada, R. I., Hudson, D., Barany, G. 1990J. Org. Chem553730Google Scholar
  3. Balbirnie, M., Grothe, R., Eisenberg, D. S. 2001Proc. Natl. Acad. Sci. U.S.A982375Google Scholar
  4. Benzinger, T. L., Gregory, D. M., Burkoth, T. S., Miller-Auer, H., Lynn, D. G., Botto, R. E., Meredith, S. C. 1998Proc. Natl. Acad. Sci. U.S.A9513407Google Scholar
  5. Booth, D. R., Sunde, M., Bellotti, V., Robinson, C. V., Hutchinson, W. L., Fraser, P. E., Hawkins, P. N., Dobson, C. M., Radford, S. E., Blake, C. C., Pepys, M. B. 1997Nature385787CrossRefGoogle Scholar
  6. Bucciantini, M., Giannoni, E., Chiti, F., Baroni, F., Formigli, L., Zurdo, J., Taddei, N., Ramponi, G., Dobson, C. M., Stefani, M. 2002Nature416507CrossRefPubMedGoogle Scholar
  7. Burke, M. G., Woscholski, R., Yaliraki, S. N. 2003Proc. Natl. Acad. Sci. U.S.A10013928Google Scholar
  8. Burkhard, P., Meier, M., Lustig, A. 2000Protein Sci92294Google Scholar
  9. Burkhard, P., Stetefeld, J., Strelkov, S. V. 2001Trends Cell Biol1182CrossRefPubMedGoogle Scholar
  10. Cerpa, R., Cohen, F. E., Kuntz, I. D. 1996Fold. Des191Google Scholar
  11. Chakrabartty, A., Baldwin, R. L. 1995Adv. Protein Chem46141Google Scholar
  12. Chiba, T., Hagihara, Y., Higurashi, T., Hasegawa, K., Naiki, H., Goto, Y. 2003J. Biol. Chem27847016Google Scholar
  13. Chiti, F., Webster, P., Taddei, N., Clark, A., Stefani, M., Ramponi, G., Dobson, C. M. 1999Proc. Natl. Acad. Sci. U.S.A963590Google Scholar
  14. Chiti, F., Taddei, N., Bucciantini, M., White, P., Ramponi, G., Dobson, C. M. 2000EMBO J191441Google Scholar
  15. Chiti, F., Taddei, N., Baroni, F., Capanni, C., Stefani, M., Ramponi, G., Dobson, C. M. 2002Nat. Struct. Biol9137Google Scholar
  16. Chiti, F., Stefani, M., Taddei, N., Ramponi, G., Dobson, C.␣M. 2003Nature424805Google Scholar
  17. Ciani, B., Hutchinson, E. G., Sessions, R. B., Woolfson, D. N. 2002J. Biol. Chem27710150Google Scholar
  18. Cohen, C., Parry, D. A. 1990Proteins71Google Scholar
  19. Cohen, A. S., Shirahama, T., Skinner, M. 1982Electron microscopy of amyloidHarris, J. R. eds. Electron microscopy of proteinsAcademic PressLondon, UKGoogle Scholar
  20. Dado, P. G. and Gellman, S. H.: 1993, J. Am. Chem. Soc. , 12609. Google Scholar
  21. Grado, W. F., Summa, C. M., Pavone, V., Nastri, F., Lombardi, A. 1999Annu. Rev. Biochem68779Google Scholar
  22. Fandrich, M., Fletcher, M. A., Dobson, C. M. 2001Nature410165Google Scholar
  23. Fezoui, Y., Hartley, D. M., Walsh, D. M., Selkoe, D. J., Osterhout, J. J., Teplow, D. B. 2000Nat. Struct. Biol71095Google Scholar
  24. Frank, S., Lustig, A., Schulthess, T., Engel, J., Kammerer, R.A. 2000J. Biol. Chem27511672Google Scholar
  25. García-Echeverría, C. 1996Bioorg. Med. Chem. Lett6229Google Scholar
  26. Hammarstrom, P., Jiang, X., Hurshman, A. R., Powers, E. T., Kelly, J. W. 2002Proc. Natl. Acad. Sci. U.S.A9916427Google Scholar
  27. Harbury, P. B., Zhang, T., Kim, P. S., Alber, T. 1993Science2621401Google Scholar
  28. Hecht, M. H., Das, A., Go, A., Bradley, L. H., Wei, Y. 2004Protein Sci131711Google Scholar
  29. Hurle, M. R., Helms, L. R., Li, L., Chan, W., Wetzel, R. 1994Proc. Natl. Acad. Sci. U.S.A915446Google Scholar
  30. Janek, K., Behlke, J., Zipper, J., Fabian, H., Georgalis, Y., Beyermann, M., Bienert, M., Krause, E. 1999Biochemistry388246Google Scholar
  31. Kammerer, R. A. 1997Matrix Biol15555Google Scholar
  32. Kammerer, R. A., Schulthess, T., Landwehr, R., Lustig, A., Engel, J., Aebi, U., Steinmetz, M. O. 1998Proc. Natl. Acad. Sci. U.S.A9513419Google Scholar
  33. Kammerer, R. A., Jaravine, V. A., Frank, S., Schulthess, T., Landwehr, R., Lustig, A., Garcia-Echeverria, C., Alexandrescu, A. T., Engel, J., Steinmetz, M. O. 2001J Biol. Chem27613685Google Scholar
  34. Kammerer, R. A., Kostrewa, D., Zurdo, J., Detken, A., Garcia-Echeverria, C., Green, J. D., Muller, S. A., Meier, B. H., Winkler, F. K., Dobson, C. M., Steinmetz, M. O. 2004Proc. Natl. Acad. Sci. U.S.A1014435Google Scholar
  35. Kelly, J. W., Balch, W. E. 2003J. Cell Biol161461Google Scholar
  36. Kohn, W. D., Hodges, R. S. 1998Trends Biotechnol16379Google Scholar
  37. Kyte, J., Doolittle, R. F. 1982J Mol. Biol157105Google Scholar
  38. Lansbury, P. T.,Jr. 1999Proc. Natl. Acad. Sci. U.S.A963342Google Scholar
  39. Liemann, S., Glockshuber, R. 1999Biochemistry383258CrossRefPubMedGoogle Scholar
  40. Lopez, P. M., Goldie, K., Zurdo, J., Lacroix, E., Dobson, C.M., Hoenger, A., Serrano, L. 2002Proc. Natl. Acad. Sci. U.S.A9916052Google Scholar
  41. Lu, M., Shu, W., Ji, H., Spek, E., Wang, L., Kallenbach, N. R. 1999J Mol. Biol288743Google Scholar
  42. Lupas, A. 1996Trends Biochem. Sci21375Google Scholar
  43. McCutchen, S. L., Lai, Z., Miroy, G. J., Kelly, J. W., Colon, W. 1995Biochemistry3413527Google Scholar
  44. Minor, D. L.,Jr., Kim, P. S. 1994Nature367660CrossRefPubMedGoogle Scholar
  45. Miranker, A. D. 2004Proc. Natl. Acad. Sci. U.S.A1014335Google Scholar
  46. Nielsen, L., Frokjaer, S., Brange, J., Uversky, V. N., Fink, A.L. 2001Biochemistry408397Google Scholar
  47. Otzen, D. E., Kristensen, O., Oliveberg, M. 2000Proc. Natl. Acad. Sci. U.S.A979907Google Scholar
  48. Perutz, M. F. 1999Trends Biochem. Sci2458Google Scholar
  49. Perutz, M. F., Johnson, T., Suzuki, M., Finch, J. T. 1994Proc. Natl. Acad. Sci. U.S.A915355Google Scholar
  50. Prusiner, S. B. 2001N. Engl. J. Med3441516Google Scholar
  51. Ramirez-Alvarado, M., Merkel, J. S., Regan, L. 2000Proc. Natl. Acad. Sci. U.S.A978979Google Scholar
  52. Richardson, J. S., Richardson, D. C. 1998Science2401648Google Scholar
  53. Rochet, J. C., Lansbury, P. T.,Jr 2000Curr. Opin. Struct. Biol1060Google Scholar
  54. Sacchettini, J. C., Kelly, J. W. 2002Nat. Rev. Drug Discov 1267Google Scholar
  55. Serpell, L. C., Sunde, M., Blake, C. C. 1997Cell Mol. Life Sci53871Google Scholar
  56. Smith, D. P., Jones, S., Serpell, L. C., Sunde, M., Radford, S. E. 2003J. Mol. Biol330943Google Scholar
  57. Stefani, M., Dobson, C. M. 2003J. Mol. Med81678Google Scholar
  58. Steinmetz, M. O., Stock, A., Schulthess, T., Landwehr, R., Lustig, A., Faix, J., Gerisch, G., Aebi, U., Kammerer, R. A. 1998EMBO J171883Google Scholar
  59. Su, J. Y., Hodges, R. S., Kay, C. M. 1994Biochemistry3315501Google Scholar
  60. Sunde, M., Serpell, L. C., Bartlam, M., Fraser, P. E., Pepys, M. B., Blake, C. C. 1997J. Mol. Biol273729Google Scholar
  61. Takahashi, Y., Ueno, A., Mihara, H. 2000Structure. Fold. Des8915Google Scholar
  62. Taylor, J. P., Hardy, J., Fischbeck, K. H. 2002Science2961991Google Scholar
  63. Tenidis, K., Waldner, M., Bernhagen, J., Fischle, W., Bergmann, M., Weber, M., Merkle, M. L., Voelter, W., Brunner, H., Kapurniotu, A. 2000J. Mol. Biol2951055Google Scholar
  64. Tjernberg, L. O., Callaway, D. J., Tjernberg, A., Hahne, S., Lilliehook, C., Terenius, L., Thyberg, J., Nordstedt, C. 1999J. Biol. Chem27412619Google Scholar
  65. Tjernberg, L., Hosia, W., Bark, N., Thyberg, J., Johansson, J. 2002aJ. Biol. Chem27743243Google Scholar
  66. Tjernberg, L. O., Tjernberg, A., Bark, N., Shi, Y., Ruzsicska, B. P., Bu, Z., Thyberg, J., Callaway, D. J. 2002bBiochem. J366343Google Scholar
  67. Villegas, V., Zurdo, J., Filimonov, V. V., Aviles, F. X., Dobson, C. M., Serrano, L. 2000Protein Sci91700Google Scholar
  68. Walsh, D. M., Hartley, D. M., Kusumoto, Y., Fezoui, Y., Condron, M. M., Lomakin, A., Benedek, G. B., Selkoe, D. J., Teplow, D. B. 1999J. Biol. Chem27425945Google Scholar
  69. West, M. W., Wang, W., Patterson, J., Mancias, J. D., Beasley, J. R., Hecht, M. H. 1999Proc. Natl. Acad. Sci. U.S.A 9611211Google Scholar
  70. Zhang, S. 2003Nat. Biotechnol211171Google Scholar
  71. Zhang, S., Rich, A. 1997Proc. Natl. Acad. Sci. U.S.A9423Google Scholar
  72. Zhu, L., Zhang, X. J., Wang, L. Y., Zhou, J. M., Perrett, S. 2003J. Mol. Biol328235Google Scholar

Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Michel O. Steinmetz
    • 1
  • Carlos García-Echeverría
    • 2
  • Richard A. Kammerer
    • 3
  1. 1.Biomolecular Research, Structural BiologyPaul Scherrer InstituteVilligen PSISwitzerland
  2. 2.Novartis Institutes for BioMedical ResearchBaselSwitzerland
  3. 3.Wellcome Trust Centre for Cell-Matrix ResearchSchool of Biological Sciences, University of ManchesterManchesterUK

Personalised recommendations