Amyloid fibrils are thread-like protein aggregates with a core region formed from repetitive arrays of β-sheets oriented parallel to the fibril axis1,2. Such structures were first recognized in clinical disorders1,3, but more recently have also been linked to a variety of non-pathogenic phenomena ranging from the transfer of genetic information to synaptic changes associated with memory4,5,6,7. The observation that many proteins can convert into similar structures in vitro has suggested that this ability is a generic feature of polypeptide chains1,8. Here we have probed the nature of the amyloid structure by monitoring hydrogen/deuterium exchange in fibrils formed from an SH3 domain9,10,11,12 using a combination of nuclear magnetic resonance spectroscopy and electrospray ionization mass spectrometry. The results reveal that under the conditions used in this study, exchange is dominated by a mechanism of dissociation and re-association that results in the recycling of molecules within the fibril population. This insight into the dynamic nature of amyloid fibrils, and the ability to determine the parameters that define this behaviour, have important implications for the design of therapeutic strategies directed against amyloid disease.
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Dobson, C. M. Protein folding and misfolding. Nature 426, 884–890 (2003)
Sunde, M. & Blake, C. The structure of amyloid fibrils by electron microscopy and X-ray diffraction. Advan. Protein Chem. 50, 123–159 (1997)
Selkoe, D. J. Folding proteins in fatal ways. Nature 426, 900–904 (2003)
Wickner, R. B. et al. Prions: proteins as genes and infectious entities. Genes Dev. 18, 470–485 (2004)
Chapman, M. R. et al. Role of Escherichia coli curli operons in directing amyloid fiber formation. Science 295, 851–855 (2002)
Kelly, J. W. & Balch, W. E. Amyloid as a natural product. J. Cell Biol. 161, 461–462 (2003)
Si, K., Lindquist, S. & Kandel, E. R. A neuronal isoform of the Aplysia CPEB has prion-like properties. Cell 115, 879–891 (2003)
Stefani, M. & Dobson, C. M. Protein aggregation and aggregate toxicity: new insights into protein folding, misfolding diseases and biological evolution. J. Mol. Med. 81, 678–699 (2003)
Jiménez, J. L. et al. Cryo-electron microscopy structure of an SH3 amyloid fibril and model of the molecular packing. EMBO J. 18, 815–821 (1999)
Zurdo, J., Guijarro, J. I., Jiménez, J. L., Saibil, H. R. & Dobson, C. M. Dependence on solution conditions of aggregation and amyloid formation by an SH3 domain. J. Mol. Biol. 311, 325–340 (2001)
Zurdo, J., Guijarro, J. I. & Dobson, C. M. Preparation and characterization of purified amyloid fibrils. J. Am. Chem. Soc. 123, 8141–8142 (2001)
Ventura, S. et al. Short amino acid stretches can mediate amyloid formation in globular proteins: the Src homology 3 (SH3) case. Proc. Natl Acad. Sci. USA 101, 7258–7263 (2004)
Englander, S. W. & Krishna, M. M. G. Hydrogen exchange. Nature Struct. Biol. 8, 741–742 (2001)
Kheterpal, I., Zhou, S., Cook, K. D. & Wetzel, R. Aβ amyloid fibrils possess a core structure highly resistant to hydrogen exchange. Proc. Natl Acad. Sci. USA 97, 13597–13601 (2000)
Hoshino, M. et al. Mapping the core of the β2-microglobulin amyloid fibril by H/D exchange. Nature Struct. Biol. 9, 332–336 (2002)
Olofsson, A., Ippel, J. H., Wijmenga, S. S., Lundgren, E. & Oehman, A. Probing solvent accessibility of transthyretin amyloid by solution NMR spectroscopy. J. Biol. Chem. 279, 5699–5707 (2004)
Petkova, A. T. et al. Self-propagating, molecular-level polymorphism in Alzheimer's β-amyloid fibrils. Science 307, 262–265 (2005)
Polverino de Laureto, P. et al. Protein aggregation and amyloid fibril formation by an SH3 domain probed by limited proteolysis. J. Mol. Biol. 334, 129–141 (2003)
Miranker, A., Robinson, C. V., Radford, S. E., Aplin, R. T. & Dobson, C. M. Detection of transient protein folding populations by mass spectrometry. Science 262, 896–900 (1993)
Yamaguchi, K. et al. Core and heterogeneity of β2-microglobulin amyloid fibrils as revealed by H/D exchange. J. Mol. Biol. 338, 559–571 (2004)
Kheterpal, I. et al. Aβ protofibrils possess a stable core structure resistant to hydrogen exchange. Biochemistry 42, 14092–14098 (2003)
Ban, T., Hamada, D., Hasegawa, K., Naiki, H. & Goto, Y. Direct observation of amyloid fibril growth monitored by thioflavin T fluorescence. J. Biol. Chem. 278, 16462–16465 (2003)
Goldsbury, C., Kistler, J., Aebi, U., Arvinte, T. & Cooper, G. J. S. Watching amyloid fibrils grow by time-lapse atomic force microscopy. J. Mol. Biol. 285, 33–39 (1999)
Hall, D. & Edskes, H. Silent prions lying in wait: a two-hit model of prion/amyloid formation and infection. J. Mol. Biol. 336, 775–786 (2004)
Hammarstroem, P., Wiseman, R. L., Powers, E. T. & Kelly, J. W. Prevention of transthyretin amyloid disease by changing protein misfolding energetics. Science 299, 713–716 (2003)
Sekijima, Y. et al. The biological and chemical basis for tissue-selective amyloid disease. Cell 121, 73–85 (2005)
Cohen, F. E. & Kelly, J. W. Therapeutic approaches to protein-misfolding diseases. Nature 426, 905–909 (2003)
Hamada, D., Yanagihara, I. & Tsumoto, K. Engineering amyloidogenicity towards the development of nanofibrillar materials. Trends Biotechnol. 22, 93–97 (2004)
MacPhee, C. E. & Woolfson, D. N. Engineered and designed peptide-based fibrous biomaterials. Curr. Opin. Solid State Mater. Sci. 8, 141–149 (2004)
Dobson, C. M. In the footsteps of alchemists. Science 304, 1261–1262 (2004)
N.C. acknowledges receipt of a Marie Curie Fellowship and a Marie Curie Reintegration Grant from the EC. G.L.C. acknowledges support from the Walters-Kundert Trust and the EPSRC. D.R.H. acknowledges receipt of an HFSP Fellowship. The research of C.V.R. is supported in part by the Royal Society, and that of C.M.D. and J.Z. by Programme Grants from the Wellcome Trust and the Leverhulme Trust.
Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
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Carulla, N., Caddy, G., Hall, D. et al. Molecular recycling within amyloid fibrils. Nature 436, 554–558 (2005) doi:10.1038/nature03986
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