Abstract
Amyloid fibrils are attractive targets for applications in biotechnology. These thin, nanoscale protein fibers are highly ordered structures that self-assemble from their component proteins or peptides. This chapter describes the use of several biophysical techniques to monitor the formation of amyloid fibrils including a common dye-binding assay, turbidity assay, and small-angle X-ray scattering. These techniques provide information about the assembly mechanism, the rate and reproducibility of assembly, as well as the size of species along the assembly pathway.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Guijarro J, Sunde M, Jones J, Campbell I, Dobson CM (1998) Amyloid fibril formation by an sh3 domain. Proc Natl Acad Sci U S A 95:4224–4228
Dobson CM (1999) Protein misfolding, evolution and disease. Trends Biochem Sci 24:329–332
Fandrich M, Fletcher M, Dobson CM (2001) Amyloid fibrils from muscle myoglobin. Nature 410:165–166
Gras SL (2007) Amyloid fibrils: from disease to design. New biomaterial applications for self-assembling cross-β fibrils. Aust J Chem 60:333–342
Nilsberth C, Westlind-Danielsson A, Eckman CB, Condron MM, Axelman K, Forsell C, Stenh C, Luthman J, Teplow DB, Younkin SG, Naslund J, Lannfelt L (2001) The ‘arctic’ APP mutation (E693G) causes Alzheimer’s disease by enhanced A-beta protofibril formation. Nat Neurosci 4:887–893
Jones L (2002) The cell biology of huntington’s disease. Oxford University Press, Oxford OX2 6DP, UK, pp 348–362
Olsen A, Jonsson A, Normark S (1989) Fibronectin binding mediated by a novel class of surface organelles on Escherichia coli. Nature 338:652–655
Saupe SJ (2000) Molecular genetics of heterokaryon incompatibility in filamentous Ascomycetes. Microbiol Mol Biol Rev 64:489–502
Balguerie A, Dos Reis S, Ritter C, Chaignepain S, Coulary-Salin B, Forge V, Bathany K, Lascu I, Scmitter J, Riek R, Saupe SJ (2003) Domain organization and structure-function relationship of the het-s prion protein of Podospora anserina. EMBO J 22:2071–2081
Barlow D, Dickinson G, Orihuela B, Kulp J III, Rittschof D, Wahl K (2010) Characterization of the adhesive plaque of the barnacle balanus amphitrite: amyloid-like nanofibrils are a major component. Langmuir 26:6549–6556
Berson JF, Theos AC, Harper DC, Tenza D, Raposo G, Marks MS (2003) Proprotein convertase cleavage liberates a fibrillogenic fragment of a resident glycoprotein to initiate melanosome biogenesis. J Cell Biol 161:461–462
Fowler DM, Koulov AV, Alory-Jost C, Marks MS, Balch WE, Kelly JW (2006) Functional amyloid formation within mammalian tissue. PLoS Biol 4:e6
Caflisch A (2006) Computational models for the prediction of polypeptide aggregation propensity. Curr Opin Chem Biol 10:437–444
Conchillo-Sole O, de Groot NS, Aviles FX, Vendrell J, Daura X, Ventura S (2007) Aggrescan: a server for the prediction and evaluation of ‘hot spots’ of aggregation in polypeptides. BMC Bioinformatics 8:65–81
Nadaud PS, Sarkar M, Wu B, MacPhee CE, Magliery TJ, Jaroniec CP (2010) Expression and purification of a recombinant amyloidogenic peptide from transthyretin for solid-state NMR spectroscopy. Protein Exp Purif 70:101–108
Tickler AK, Clippingdale AB, Wade JD (2004) Amyloid-beta as a ‘difficult sequence’ in solid phase peptide synthesis. Protein Pept Lett 11:377–384
Gustavsson A, Engstrom U, Westermark P (1991) Normal transthyretin and synthetic transthyretin fragments form amyloid-like fibrils in vitro. Biochem Biophys Res Commun 175:1159–1164
Gras SL, Tickler AK, Squires AM, Devlin GL, Horton MA, Dobson CM, MacPhee CE (2008) Functionalised amyloid fibrils for roles in cell adhesion. Biomaterials 3:22–30
de La Paz ML, Goldie K, Zurdo J, Lacroix E, Dobson CM, Hoenger A, Serrano L (2002) De novo designed peptide-based amyloid fibrils. Proc Natl Acad Sci U S A 99:16052–16057
Garvey M, Gras S, Meehan S, Meade S, Carver J, Gerrard J (2009) Protein nanofibres of defined morphology prepared from mixtures of crude crystallins. Int J Nanotechnol 6:258–273
Wetzel R (2006) Kinetics and thermodynamics of amyloid fibril assembly. Acc Chem Res 39:671–679
Nilsson M (2004) Techniques to study amyloid fibril formation in vitro. Methods 34:151–160
Naiki H, Higuchi K, Hosokawa M, Takeda T (1989) Fluorometric-determination of amyloid fibrils in vitro using the fluorescent dye. Thioflavine-T. Anal Biochem 177:244–249
LeVine H (1993) Thioflavine-T interaction with synthetic Alzheimer’s disease beta-amyloid peptides - detection of amyloid aggregation in solution. Protein Sci 2:404–410
Krebs MRH, Bromley EHC, Donald AM (2005) The binding of Thioflavin-T to amyloid fibrils: localisation and implications. J Struct Biol 149:30–37
Groenning M, Norrman M, Flink JM, van de Weert M, Bukrinsky JT, Schluckebier G, Frokjaer S (2007) Binding mode of Thioflavin-T in insulin amyloid fibrils. J Struct Biol 159:483–497
Kitts CC, Bout DAV (2009) Near-field scanning optical microscopy measurements of fluorescent molecular probes binding to insulin amyloid fibrils. J Phys Chem B 113:12090–12095
Wolfe LS, Calabrese MF, Nath A, Blaho DV, Miranker AD, Xiong Y (2010) Protein-induced photophysical changes to the amyloid indicator dye Thioflavin-T. Proc Natl Acad Sci U S A 107:16863–16868
Biancalana M, Koide S (2010) Molecular mechanism of Thioflavin-T binding to amyloid fibrils. Biochim Biophys Acta 1804:1405–1412
Svergun D, Koch M (2003) Small-angle scattering studies of biological macromolecules. Rep Prog Phys 66:1735–1782
Putnam C, Hammel M, Hura G, Tainer J (2007) X-ray solution scattering (SAXS) combined with crystallography and computation: defining accurate macromolecular structures, conformations and assemblies in solution. Q Rev Biophys 40:191–285
Konarev P, Petoukhov M, Volkov V, Svergun D (2006) Atsas 2.1, a program package for small-angle scattering data analysis. J Appl Crystallogr 39:277–286
Chen S, Wetzel R (2001) Solubilization and disaggregation of polyglutamine peptides. Protein Sci 10:887–891
Fezoui Y, Hartley DM, Harper JD, Khurana R, Walsh DM, Condron MM, Selkoe DJ, Lansbury PTJ, Fink AL, Teplow DB (2000) An improved method of preparing the amyloid beta-protein for fibrillogenesis and neurotoxicity experiments. Amyloid 7:166–178
Lashuel HA, Hartley DM, Petre BM, Wall JS, Simon MN, Walz T, Lansbury PTJ (2003) Mixtures of wild-type and a pathogenic (E22G) form of Aβ40 in vitro accumulate protofibrils, including amyloid pores. J Mol Biol 332:795–808
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Sawyer, E.B., Gras, S.L. (2013). Self-Assembling Nanomaterials: Monitoring the Formation of Amyloid Fibrils, with a Focus on Small-Angle X-Ray Scattering. In: Gerrard, J. (eds) Protein Nanotechnology. Methods in Molecular Biology, vol 996. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-354-1_5
Download citation
DOI: https://doi.org/10.1007/978-1-62703-354-1_5
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-353-4
Online ISBN: 978-1-62703-354-1
eBook Packages: Springer Protocols