Abstract
Amyloid formation has been implicated in the pathology of over 20 human diseases, but the rational design of amyloid inhibitors is hampered by a lack of structural information about amyloid–inhibitor complexes. We use isotope labelling and two-dimensional infrared spectroscopy to obtain a residue-specific structure for the complex of human amylin (the peptide responsible for islet amyloid formation in type 2 diabetes) with a known inhibitor (rat amylin). Based on its sequence, rat amylin should block formation of the C-terminal β-sheet, but at 8 h after mixing, rat amylin blocks the N-terminal β-sheet instead. At 24 h after mixing, rat amylin blocks neither β-sheet and forms its own β-sheet, most probably on the outside of the human fibrils. This is striking, because rat amylin is natively disordered and not previously known to form amyloid β-sheets. The results show that even seemingly intuitive inhibitors may function by unforeseen and complex structural processes.
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Acknowledgements
Support for this research was provided by the National Institutes of Health (grants DK79895 to M.T.Z., GM078114 to D.P.R. and DK088184 to J.J.d.P.) and the National Science Foundation (CRC grant CHE 0832584 to M.T.Z and D.P.R.). The authors are grateful to R. Tycko for providing the coordinates for his structural model of human amylin fibrils.
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C.T.M., D.P.R. and M.T.Z. designed the research. C.T.M. performed the 2D IR and electron microscopy measurements. P.M., A.W.M. and P.C. synthesized and purified peptides. P.M and P.C performed the thioflavin-T fluorescence measurements. C.C., S.S. and J.J.d.P. designed and performed the molecular dynamics simulations. C.T.M. and M.T.Z. analysed data. C.T.M., D.P.R. and M.T.Z. wrote the manuscript and coordinated contributions by other authors.
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Middleton, C., Marek, P., Cao, P. et al. Two-dimensional infrared spectroscopy reveals the complex behaviour of an amyloid fibril inhibitor. Nature Chem 4, 355–360 (2012). https://doi.org/10.1038/nchem.1293
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DOI: https://doi.org/10.1038/nchem.1293
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