Abstract
Amyloid fibrils are a highly ordered and robust aggregated form of protein structure in which the protein components are arranged in long fibrillar arrays comprised of β-sheet. Because of these properties, along with their biocompatibility, amyloid fibrils have attracted much research attention as bionanomaterials, for example as template structures (in some cases following modification) that can be used as biosensors, encapsulators, and biomimetic materials. To use amyloid fibrils for such a range of applications will require them to be obtained relatively easily in large quantities. In this chapter, we describe methods for isolating crystallin and casein proteins from readily available sources that contain abundant protein, i.e., the eye lens and milk, respectively, and the subsequent conversion of these proteins into amyloid fibrils.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chiti F, Dobson CM (2006) Protein misfolding, functional amyloid, and human disease. Annu Rev Biochem 75:333–366
Ecroyd H, Carver JA (2008) Unraveling the mysteries of protein folding and misfolding. IUBMB Life 60:769–774
Jiménez JL, Nettleton EJ, Bouchard M, Robinson CV, Dobson CM, Saibil HR (2002) The protofilament structure of insulin amyloid fibrils. Proc Natl Acad Sci USA 99:9196–9201
Fowler DM, Koulov AV, Balch WE, Kelly JW (2007) Functional amyloid—from bacteria to humans. Trends Biochem Sci 32:217–224
Garvey M, Gras SL, Meehan S, Meade SJ, Carver JA, Gerrard JA (2009) Protein nanofibres of defined morphology prepared from mixtures of crude crystallins. Int J Nanotech 6:258–273
Gras SL (2007) Amyloid fibrils: From disease to design. New biomaterial applications for self-assembling cross beta-fibrils. Aust J Chem 60:333–342
Cherny I, Gazit E (2008) Amyloids: not only pathological agents but also ordered nanomaterials. Angew Chem Int Ed Engl 47:4062–4069
Bloemendal H, de Jong W, Jaenicke R, Lubsen NH, Slingsby C, Tardieu A (2004) Ageing and vision: structure, stability and function of lens crystallins. Prog Biophys Mol Biol 86:407–485
Meehan S, Berry Y, Luisi B, Dobson CM, Carver JA, MacPhee CE (2004) Amyloid fibril formation by lens crystallin proteins and its implications for cataract formation. J Biol Chem 279:3413–3419
Meehan S, Knowles TP, Baldwin AJ, Smith JF, Squires AM, Clements P, Treweek TM, Ecroyd H, Tartaglia GG, Vendruscolo M, Macphee CE, Dobson CM, Carver JA (2007) Characterisation of amyloid fibril formation by small heat-shock chaperone proteins human alphaA-, alphaB- and R120G alphaB-crystallins. J Mol Biol 372:470–484
Horwitz J, Huang QL, Ding L, Bova MP (1998) Lens alpha-crystallin: chaperone-like properties. Methods Enzymol 290:365–383
Thorn DC, Ecroyd H, Carver JA (2009) The two-faced nature of milk casein proteins: amyloid fibril formation and chaperone-like activity. Aust J Dairy Technol 64:36–40
Thorn DC, Ecroyd H, Sunde M, Poon S, Carver JA (2008) Amyloid fibril formation by bovine milk alphaS2-casein occurs under physiological conditions yet is prevented by its natural counterpart, alphaS1-casein. Biochemistry 47:3926–3936
Thorn DC, Meehan S, Sunde M, Rekas A, Gras SL, MacPhee CE, Dobson CM, Wilson MR, Carver JA (2005) Amyloid fibril formation by bovine milk kappa-casein and its inhibition by the molecular chaperones alpha(S)- and beta-casein. Biochemistry 44:17027–17036
Morgan PE, Treweek TM, Lindner RA, Price WE, Carver JA (2005) Casein proteins as molecular chaperones. J Agric Food Chem 53:2670–2683
Ecroyd H, Thorn DC, Liu Y, Carver JA (2010) The dissociated form of kappa-casein is the precursor to its amyloid fibril formation. Biochem J 429:251–260
Ecroyd H, Koudelka T, Thorn DC, Williams DM, Devlin G, Hoffmann P, Carver JA (2008) Dissociation from the oligomeric state is the rate-limiting step in amyloid fibril formation by kappa-casein. J Biol Chem 283:9012–9022
Carver JA, Duggan PJ, Ecroyd H, Liu Y, Meyer AG, Tranberg CE (2010) Carboxymethylated-kappa-casein: a convenient tool for the identification of polyphenolic inhibitors of amyloid fibril formation. Bioorg Med Chem 18:222–228
Acknowledgments
This work was supported by grants from the Australian Research Council and Dairy Australia (DA). D.T. was supported by a postgraduate scholarship from DA and M.G. was supported by a postgraduate scholarship from Crop and Food Research New Zealand funded by the Foundation for Research Science and Technology.
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
Ecroyd, H., Garvey, M., Thorn, D.C., Gerrard, J.A., Carver, J.A. (2013). Amyloid Fibrils from Readily Available Sources: Milk Casein and Lens Crystallin Proteins. 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_6
Download citation
DOI: https://doi.org/10.1007/978-1-62703-354-1_6
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