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An infrared spectroscopy approach to follow β-sheet formation in peptide amyloid assemblies

Abstract

Amyloidogenic peptides and proteins play a crucial role in a variety of neurodegenerative disorders such as Alzheimer's and Parkinson's disease. These proteins undergo a spontaneous transition from a soluble, often partially folded form, into insoluble amyloid fibrils that are rich in β-sheets. Increasing evidence suggests that highly dynamic, polydisperse folding intermediates, which occur during fibril formation, are the toxic species in the amyloid-related diseases. Traditional condensed-phase methods are of limited use for characterizing these states because they typically only provide ensemble averages rather than information about individual oligomers. Here we report the first direct secondary-structure analysis of individual amyloid intermediates using a combination of ion mobility spectrometry–mass spectrometry and gas-phase infrared spectroscopy. Our data reveal that oligomers of the fibril-forming peptide segments VEALYL and YVEALL, which consist of 4–9 peptide strands, can contain a significant amount of β-sheet. In addition, our data show that the more-extended variants of each oligomer generally exhibit increased β-sheet content.

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Figure 1: Structural analysis of VEALYL oligomers and fibrils in the condensed phase and in the gas phase.
Figure 2: TEM images of VEALYL sequence variants.
Figure 3: IMS and conformer-selected infrared spectroscopy of YVEALL oligomers.
Figure 4: Correlation between increase in CCS and β-sheet fraction.

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Acknowledgements

The authors thank S. Huhmann for the help during synthesis, L. Urner for fruitful discussion and R. Schlögl for proofreading the manuscript. B. Koksch and M. Villinger are gratefully acknowledged for providing the peptide synthesis facilities and EM infrastructure. M.T.B. acknowledges the Alexander von Humboldt-Foundation and the National Science Foundation for support under grant CHE-1301032.

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J.S., W.H., M.T.B., G.v.H. and K.P. conceived and designed the experiments; J.S., W.H. and S.W. performed the experiments: X.H., S.G. and W.S. supported the experiments; J.S. and W.H. analysed data; all the authors co-wrote the paper. J.S. and W.H. contributed equally to this work.

Corresponding authors

Correspondence to Gert von Helden or Kevin Pagel.

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The authors declare no competing financial interests.

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Seo, J., Hoffmann, W., Warnke, S. et al. An infrared spectroscopy approach to follow β-sheet formation in peptide amyloid assemblies. Nature Chem 9, 39–44 (2017). https://doi.org/10.1038/nchem.2615

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