Abstract
Twenty-five proteins are known to form amyloid fibrils in vivo in association with disease (Westermark et al., Amyloid 12:1–4, 2005). However, the fundamental ability of a protein to form amyloid-like fibrils is far more widespread than in just the proteins associated with disease, and indeed this property can provide insight into the basic thermodynamics of folding and misfolding pathways. But how does one determine whether a protein has formed amyloid-like fibrils? In this chapter, we cover the basic steps toward defining the amyloid-like properties of a protein and how to measure the kinetics of fibrillization. We describe several basic tests for aggregation and the binding to two classic amyloid-reactive dyes, Congo Red, and thioflavin T, which are key indicators to the presence of fibrils.
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Virchow R. (1854) Ueber eine im gehirn und ruckenmark des menschen aufgefunde substanz mit der chemishen reaction der cellulose. Virchows Arch Path Anat 6, 135–138.
Divry P., and Florkin M. (1927) Sur les proprietes optiques de l’amyloide.
Comptes Rendus de la Societe de Biologie 97, 1808–1810.
Missmahl H. P., and Hartwig M. (1953) Polarisationsoptische untersuchungen an der amyloidsubstanz. Virchows Archiv 324, 489–508.
Westermark P., Benson M. D., Buxbaum J. N., Cohen A. S., Frangione B., Ikeda S.-I., Masters C. L., Merlini G., Saraiva M. J., and Sipe J. D. (2005) Amyloid: Toward terminology clarification report from the nomenclature committee of the international society of amyloidosis. Amyloid 12, 1–4.
Cohen A. S., and Calkins E. (1959) Electron microscopic observations on a fibrous component in amyloid of diverse origins. Nature 183, 1202–1203.
Chiti F., and Dobson C. M. (2006) Protein misfolding, functional amyloid, and human disease. Ann. Rev. Biochem. 75, 333–366.
Sipe J. D., and Cohen A. S. (2000) Review: History of the amyloid fibril. J. Struct. Biol. 130, 88–98.
Serpell L. C. (2000) Alzheimer’s amyloid fibrils: Structure and assembly. Biochim. Biophys. Acta 1502, 16–30.
Sunde M., and Blake C. (1997) The structure of amyloid fibrils by electron microscopy and x-ray diffraction. Adv. Protein Chem. 50, 123–159.
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., and Pepys M. B. (1997) Instability, unfolding and aggregation of human lysozyme variants underlying amyloid fibrillogenesis. Nature 385, 787–793.
Fandrich M., Fletcher M. A., and Dobson C. M. (2001) Amyloid fibrils from muscle myoglobin. Nature 410, 165–166.
Chapman M. R., Robinson L. S., Pinkner J. S., Roth R., Heuser J., Hammar M., Normark S., and Hultgren S. J. (2002) Role of Escherichia coli curli operons in directing amyloid fiber formation. Science 295, 851–855.
Hatters D. M., Zhong N., Rutenber E., and Weisgraber K. H. (2006) Amino-terminal domain stability mediates apolipoprotein E aggregation into neurotoxic fibrils. J. Mol. Biol. 361, 932–944.
Klunk W. E., Jacob R. F., and Mason R. P. (1999) Quantifying amyloid β-peptide (Aβ) aggregation using the congo red-Aβ (CR-Aβ) spectrophotometric assay. Anal Biochem 266, 66–76.
Vassar P. S., and Culling C. F. (1959) Fluorescent stains, with special reference to amyloid and connective tissues. Arch. Pathol. 68, 487–498.
Kelenyi G. (1967) On the histochemistry of azo group-free thiazole dyes. J Histochem. Cytochem. 15, 172–180.
LeVine H., 3rd. (1993) Thioflavine T interaction with synthetic alzheimer’s disease β-amyloid peptides: Detection of amyloid aggregation in solution. Protein Sci. 2, 404–410.
Lowry O. H., Rosebrough N. J., Farr A. L., and Randall R. J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275.
Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., and Klenk D. C. (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150, 76–85.
True H. L., and Lindquist S. L. (2000) A yeast prion provides a mechanism for genetic variation and phenotypic diversity. Nature 407, 477–483.
Hatters D. M., MacPhee C. E., Lawrence L. J., Sawyer W. H., and Howlett G. J. (2000) Human apolipoprotein C-II forms twisted amyloid ribbons and closed loops. Biochemistry 39, 8276–8283.
Bradford M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72, 248–254.
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Hatters, D.M., Griffin, M.D.W. (2011). Diagnostics for Amyloid Fibril Formation: Where to Begin?. In: Hill, A., Barnham, K., Bottomley, S., Cappai, R. (eds) Protein Folding, Misfolding, and Disease. Methods in Molecular Biology, vol 752. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-223-0_8
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DOI: https://doi.org/10.1007/978-1-60327-223-0_8
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