Abstract
Solid-state NMR spectroscopy (SSNMR) is an established and invaluable tool for the study of amyloid fibril structure with atomic-level detail. Optimization of the homogeneity and concentration of fibrils enhances the resolution and sensitivity of SSNMR spectra. Here, we present a fibrillization and fibril processing protocol, starting from purified monomeric α-synuclein, that enables the collection of high-resolution SSNMR spectra suitable for site-specific structural analysis. This protocol does not rely on any special features of α-synuclein and should be generalizable to any other amyloid protein.
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Acknowledgements
This work was supported by NIH grants R01-GM073770 and P50-NS053488. M.D.T. and A.M.B. were supported by NIH Training Grant PHS 5T32 GM008276 and J.M.C. was supported by a National Science Foundation Graduate Research Fellowship.
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Tuttle, M.D., Courtney, J.M., Barclay, A.M., Rienstra, C.M. (2016). Preparation of Amyloid Fibrils for Magic-Angle Spinning Solid-State NMR Spectroscopy. In: Eliezer, D. (eds) Protein Amyloid Aggregation. Methods in Molecular Biology, vol 1345. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2978-8_11
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DOI: https://doi.org/10.1007/978-1-4939-2978-8_11
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2977-1
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