Abstract
The pathogenesis of a large number of diseases, including Alzheimer’s Disease, Parkinson’s Disease, and Creutzfeldt–Jakob Disease (CJD), is associated with protein aggregation and the formation of amyloid, fibrillar deposits. Peptide fragments of amyloid-forming proteins have been found to form fibrils in their own right and have become important tools for unlocking the mechanism of amyloid fibril formation and the pathogenesis of amyloid diseases. The synthesis and purification of peptide sequences derived from amyloid fibril-forming proteins can be extremely challenging. The synthesis may not proceed well, generating a very low quality crude product which can be difficult to purify. Even clean crude peptides can be difficult to purify, as they are often insoluble or form fibrils rapidly in solution. This chapter presents methods to recognise and to overcome the difficulties associated with the synthesis, and purification of fibril-forming peptides, illustrating the points with three synthetic examples.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Merrifield, R. B. (1963) Solid Phase Synthesis I. The Synthesis of a Tetrapeptide. J. Amer. Chem. Soc. 85, 2149–2154.
Kent, S. B., Mitchell, A. R., Engelhard, M. and Merrifield, R. B.(1979) Mechanisms and prevention of trifluoroacetylation in solid-phase peptide synthesis. Proc. Natl. Acad. Sci. U S A. 76(5), 2180–2184.
Sarin, V. K., Kent, S. B., Tam, J. P. and Merrifield, R. B. (1981) Quantitative monitoring of solid-phase peptide synthesis by the ninhydrin reaction Anal. Biochem. 117(1), 147–157.
Brown, E., Sheppard, R. C. and Williams, B. J. (1983) Peptide Synthesis. Part 5. Solid-phase Synthesis of [15-Leucine] Little Gastrin. J. Chem. Soc. Perkin Trans. I, 1161–1167.
Sheppard, R. C. (1986) Modern methods of solid-phase peptide synthesis. Science Tools 33, 9–16.
Schnolzer, M., Alewood, P., Jones, A., Alewood, D. and Kent, S. B. H. (1992) In situ neutralization in Boc-chemistry solid phase peptide synthesis. Int. J. Peptide Protein Res. 40, 180–193.
Alberico, F. and Carpino, L. A. (1997) Coupling reagents and activation Methods Enzymol. 289, 104–126.
Clark-Lewis, I., Aebersold, R., Ziltener, H., Schrader, J. W., Hood, L. E. and Kent, S. B. H. (1986) Automated Chemical Synthesis of a Protein Growth Factor for Hemopoietic Cells, Interleukin-3. Science 231, 134–139.
Scanlon, D. B., Eefting, M. A., Lloyd, C. J., Burgess, A. W. and Simpson, R. J. (1987) Synthesis of Biologically Active Transforming Growth Factor-alpha by Fluorenylmethoxycarbonyl Solid Phase Peptide Chemistry. J. Chem. Soc. Chem. Commun. 516–518.
Dawson, P. E., Muir, T. W., Clark-Lewis, I. and Kent, S. B. (1994) Synthesis of proteins by native chemical ligation. Science 266, 776–779.
Yamamoto, N., Tanabe, Y., Okamoto, R., Dawson, P.E. and Kajihara, Y. (2008) Chemical Synthesis of a Glycoprotein Having an Intact Human Complex-Type Sialyloligosaccharide under the Boc and Fmoc Synthetic Strategies J. Am. Chem. Soc. 130(2), 501–510.
Macmillan, D. (2006) Protein Synthesis: Evolving Strategies for Protein Synthesis Converge on Native Chemical Ligation Angew. Chem. Int. Ed. 45, 7668–7672.
Tickler, A. K.,. Clippingdale, A. B. and Wade, J. D. (2004) Amyloid-β as a “Difficult Sequence” in Solid Phase Peptide Synthesis. Protein & Peptide Letters 11(4), 377–384.
Mutter, M., Nefzi, A., Sato, T., Sun, X., Wahl, F., and Wohr, T. (1995) Pseudo-prolines for accessing “inaccessible” peptides Peptide Research 8(3), 145–153.
Simmonds, R. G. (1996) Use of the Hmb backbone-protecting group in the synthesis of difficult sequences. Int. J. Pept. Protein Res. 47(1–2), 36–41.
Tickler, A. K., Barrow, C. J. and Wade, J. D. (2001) Improved Preparation of Amyloid-Peptides Using DBU as N-Fmoc Deprotection Reagent J. Peptide Sci. 7, 488–494.
Sohma, Y. and Kiso, Y. (2006) “Click peptides”-chemical biology-oriented synthesis of Alzheimer’s disease-related amyloid beta peptide (abeta) analogues based on the “O-acyl isopeptide method”. Chembiochem. 7(10), 1549–1557.
Taniguchi, A., Sohma, Y., Hirayama, Y., Mukai, H., Kimura, T., Hayashi, Y., Matsuzaki, K., Kiso, Y. (2009) “Click peptide”: pH-triggered in situ production and aggregation of monomer Abeta1-42. Chembiochem. 10(4), 710–715.
Howlett, G. J., and Moore, K. J. (2006) Untangling the role of amyloid in atherosclerosis Current Opinion in Lipidology 17, 541–547.
Wilson, L. M., Mok, Y. F., Binger, K. J., Griffin, M. D., Mertens, H. D., Lin, F., Wade, J. D., Gooley, P. R. and Howlett, G. J. (2007) A structural core within apolipoprotein C-II amyloid fibrils identified using hydrogen exchange and proteolysis. J. Mol. Biol. 366(5), 1639–1651.
Van Nostrand, W.E., Davis-Salinas, J. and Saporito-Irwin, S. M. (1996) Amyloid beta-protein induces the cerebrovascular cellular pathology of Alzheimer’s disease and related disorders Ann. N Y. Acad. Sci. 777, 297–302.
Ball, H. L. and Mascagni, P. (1996) Chemical protein synthesis and purification: a methodology. Int. J. Peptide Protein Res. 48,31–47.
Bonetto, V., Massignan, T., Chiesa, R., Morbin, M., Mazzoleni, G., Diomede, L., Angeretti, N., Colombo, L., Forloni, G., Tagliavini, F., and Salmona, M. (2002) Synthetic miniprion PrP106 J. Biol. Chem. 277, 31327–31334.
Bahadi, R., Farrelly, P. V., Kenna, B. L., Kourie, J. I., Tagliavini, F., Forloni, G. and Salmona, M. (2003) PrP(82–146) homologous to a 7-kDa fragment in Channels formed with a mutant prion protein diseased brain of GSS patients Am. J. Physiol. Cell. Physiol. 285, 862–872.
Acknowledgements
We thank Ms Keyla Perez for her advice with the Vydac C4 preparative column technology and peptide purifications performed at 60°C.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Scanlon, D.B., Karas, J.A. (2011). Synthesis of Peptide Sequences Derived from Fibril-Forming Proteins. 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_3
Download citation
DOI: https://doi.org/10.1007/978-1-60327-223-0_3
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-221-6
Online ISBN: 978-1-60327-223-0
eBook Packages: Springer Protocols