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In Vitro Preparation of Prefibrillar Intermediates of Amyloid-β and α Synuclein

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Amyloid Proteins

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 299))

Abstract

Elucidating the structural properties of early intermediates (protofibrils) on the fibril formation pathway of Aβ and α-synuclein, the structural relationship among the different intermediates and their relationship to the structure of the amyloid fibrils is critical for understanding the roles of amyloid fibril formation in the pathogenesis of Alzheimer's and Parkinson's diseases. In this chapter we discuss several methods, developed by different laboratories, that enable the preparation and stabilization of amyloid-β and α-synuclein protofibrillar species of defined morphologies for biochemical, biophysical and toxicity studies.

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References

  1. Kelly, J. W. (1998) The alternative conformations of amyloidogenic proteins and their multi-step assembly pathways. Curr. Opin. Struct. Biol. 8(1), 101–106.

    Article  PubMed  CAS  Google Scholar 

  2. Rochet, J. and Lansbury, P. T. Jr. (2000) Amyloid fibrillogenesis: themes and variations. Curr. Opin. Struct. Biol. 10(1), 60–68.

    Article  PubMed  CAS  Google Scholar 

  3. Golberg, M. S. and Lansbury, P.T. Jr. (2000) Is there a cause-and-effect relationship between alpha-synuclein fibrillization and Parkinson' disease? Nat. Cell Biol. 2(7), E115–E119.

    Article  Google Scholar 

  4. Caughey, B. and Lansbury, P. T. (2003) Protofibrils, pores, fibrils, and neurodegeneration: separating the responsible protein aggregates from the innocent bystanders. Annu. Rev. Neurosci. 26, 267–298.

    Article  PubMed  CAS  Google Scholar 

  5. Volles, M. J. and Lansbury, P. T. Jr. (2003) Zeroing in on the pathogenic form of alpha-synuclein and its mechanism of neurotoxicity in Parkinson' disease. Biochemistry 42(26), 7871–7878.

    Article  PubMed  CAS  Google Scholar 

  6. Lashuel, H., et al. (2002) alpha-Synuclein, especially the Parkinson's disease-associated mutants, forms pore-like annular and tubular protofibrils. J. Mol. Biol. 322(5), 1089.

    Article  PubMed  CAS  Google Scholar 

  7. Lashuel, H. A., et al. (2002) Neurodegenerative disease: amyloid pores from pathogenic mutations. Nature 418(6895), 291.

    Article  PubMed  CAS  Google Scholar 

  8. Ding, T. T., et al. (2002) Annular alpha-synuclein protofibrils are produced when spherical protofibrils are incubated in solution or bound to brain-derived membranes. Biochemistry 41(32), 10209–10217.

    Article  PubMed  CAS  Google Scholar 

  9. Conway, K. A., et al. (2000) Acceleration of oligomerization, not fibrillization, is a shared property of both alpha-synuclein mutations linked to early-onset Parkinson' disease: implications for pathogenesis and therapy. Proc. Natl. Acad. Sci. USA 97(2), 571–576.

    Article  PubMed  CAS  Google Scholar 

  10. Harper, J. D., et al. (1999) Assembly of A beta amyloid protofibrils: an in vitro model for a possible early event in Alzheimer' disease. Biochemistry 38(28), 8972–8980.

    Article  PubMed  CAS  Google Scholar 

  11. Harper, J. D., Lieber, M., and Lansbury, P. T. Jr. (1997) Atomic force microscopic imaging of seeded fibril formation and fibril branching by the Alzheimer' disease amyloid-beta protein. Chem. Biol. 4(12), 951–959.

    Article  PubMed  CAS  Google Scholar 

  12. Lashuel, H., et al. (2003) Mixtures of wild-type and “Arctic” Abeta40 in vitro accumulate protofibrils, including amyloid pores. J. Mol. Biol. 332(4), 795–808.

    Article  PubMed  CAS  Google Scholar 

  13. Pollanen, M. S., Dickson, D. W., and Bergeron, (1993) Pathology and biology of the Lewy body. J. Neuropathol. Exp. Neurol. 52(3), 183–191.

    Article  PubMed  CAS  Google Scholar 

  14. Forno, L. S. (1996) Neuropathology of Parkinson' disease. J. Neuropathol. Exp. Neurol. 55(3), 259–272.

    Article  PubMed  CAS  Google Scholar 

  15. Selkoe, D. J. (1997) Alzheimer' disease: genotypes, phenotypes, and treatments. Science 275(5300), 630–631.

    Article  PubMed  CAS  Google Scholar 

  16. .Castano, E. M. and Frangione, B. (1988) Biology of disease human amyloidosis, Alzheimer disease and related disorders. Lab. Invest. 58(2), 122–132.

    PubMed  CAS  Google Scholar 

  17. Selkoe, D. J. (2000) Toward a comprehensive theory for Alzheimer's disease. Hypothesis: Alzheimer%s disease is caused by the cerebral accumulation and cytotoxicity of amyloid beta-protein. Ann. NY Acad. Sci. 924, 17–25.

    Article  PubMed  CAS  Google Scholar 

  18. Wang, R., et al. (1996) The profile of soluble amyloid beta protein in cultured cell media. Detection and quantification of amyloid beta protein and variants by immunoprecipitation-mass spectrometry. J. Biol. Chem. 271(50), 31894–31902.

    Article  PubMed  CAS  Google Scholar 

  19. Janus, C., et al. (2001) New developments in animal models of Alzheimer's disease. Curr. Neurol. Neurosci. Rep. 1(5), 451–457.

    Article  PubMed  CAS  Google Scholar 

  20. Lichtenthaler, S. F., et al. (1997) Mutations in the transmembrane domain of APP altering gamma-secretase specificity. Biochemistry 36(49), 15396–15403.

    Article  PubMed  CAS  Google Scholar 

  21. Janus, C., et al. (2000) A beta peptide immunization reduces behavioural impairment and plaques in a model of Alzheimer' disease. Nature 408(6815), 979–982.

    Article  PubMed  CAS  Google Scholar 

  22. Morgan, D., et al. (2000) A beta peptide vaccination prevents memory loss in an animal model of Alzheimer' disease. Nature 408(6815), 982–985.

    Article  PubMed  CAS  Google Scholar 

  23. Nilsberth, C., et al. (2001) The Arctic' APP mutation (E693G) causes Alzheimer' disease by enhanced Abeta protofibril formation. Nat. Neurosci. 4(9), 887–893.

    Article  PubMed  CAS  Google Scholar 

  24. Hoshi, M., et al. (2003) Spherical aggregates of beta-amyloid (amylospheroid) show high neurotoxicity and activate tau protein kinase I/glycogen synthase kinase-3beta. Proc. Natl. Acad. Sci. USA 100(11), 6370–6375.

    Article  PubMed  CAS  Google Scholar 

  25. Lambert, M. P., et al. (2001) Vaccination with soluble Abeta oligomers generates toxicity-neutralizing antibodies. J. Neurochem. 79(3), 595–605.

    Article  PubMed  CAS  Google Scholar 

  26. Stine, W. B. Jr., et al. (2003) In vitro characterization of conditions for amyloidbeta peptide oligomerization and fibrillogenesis. J. Biol. Chem. 278(13), 11612–11622.

    Article  PubMed  CAS  Google Scholar 

  27. Dahlgren, K. N., et al. (2002) Oligomeric and fibrillar species of amyloid-beta peptides differentially affect neuronal viability. J. Biol. Chem. 277,32046–32053.

    Article  PubMed  CAS  Google Scholar 

  28. Lambert, M. P., et al. (1998) Diffusible, nonfibrillar ligands derived from Abeta1-42 are potent central nervous system neurotoxins. Proc. Natl. Acad. Sci. USA 95(11), 6448–6453.

    Article  PubMed  CAS  Google Scholar 

  29. Klein, W. L. (2002) ADDLs & protofibrils-the missing links? Neurobiol. Aging 23(2), 231–235.

    Article  PubMed  CAS  Google Scholar 

  30. Klein, W. L., Krafft, G. A., and Finch, E. (2001) Targeting small Abeta oligomers: the solution to an Alzheimer's disease conundrum? Trends Neurosci. 24(4), 219–224.

    Article  PubMed  CAS  Google Scholar 

  31. Wang, H. W., et al. (2002) Soluble oligomers of beta amyloid (1–42) inhibit longterm potentiation but not long-term depression in rat dentate gyrus. Brain Res. 924(2), 133–140.

    Article  PubMed  CAS  Google Scholar 

  32. Harper, J. D., et al. (1997) Observation of metastable Abeta amyloid protofibrils by atomic force microscopy. Chem. Biol. 4(2), 119–125.

    Article  PubMed  CAS  Google Scholar 

  33. Walsh, D. M., et al. (1997) Amyloid beta-protein fibrillogenesis. Detection of a protofibrillar intermediate. J. Biol. Chem. 272(35), 22364–22372.

    Article  PubMed  CAS  Google Scholar 

  34. Walsh, D. M., et al. (1999) Amyloid beta-protein fibrillogenesis. Structure and biological activity of protofibrillar intermediates. J. Biol. Chem. 274(36), 25945–25952.

    Article  PubMed  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Center for Neurologic Diseases, Brigham and Women's Hospital and Department of Neurology, Harvard Medical School, Cambridge, MA

    Hilal A. Lashuel

  2. Departament de Quimica Organica, Universitat de Barcelona, Barcelona, Spain

    Dolors Grillo-Bosch

Authors
  1. Hilal A. Lashuel
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  2. Dolors Grillo-Bosch
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Editor information

Editors and Affiliations

  1. Departments of Psychiatry and Pathology, New York University School of Medicine, New York, NY

    Einar M. Sigurdsson

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© 2005 Humana Press Inc.

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Lashuel, H.A., Grillo-Bosch, D. (2005). In Vitro Preparation of Prefibrillar Intermediates of Amyloid-β and α Synuclein. In: Sigurdsson, E.M. (eds) Amyloid Proteins. Methods in Molecular Biology™, vol 299. Humana Press. https://doi.org/10.1385/1-59259-874-9:019

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  • DOI: https://doi.org/10.1385/1-59259-874-9:019

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-337-4

  • Online ISBN: 978-1-59259-874-8

  • eBook Packages: Springer Protocols

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