Abstract
The formation of amyloid fibrils is a central phenomenon in the progressive pathology of many neurodegenerative diseases, as well as in the fabrication of functional materials. Several different molecular processes acting in concert are responsible for the formation of amyloid fibrils from monomeric protein in solution. Here, we describe a method to determine which microscopic processes drive the overall formation of fibrils by using chemical kinetics in combination with systematic experimental datasets analysed in a global manner. We outline general concepts for obtaining suitable kinetic data and detail the key stages of data analysis, from quality control to the verification of a specific mechanism of aggregation.
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Acknowledgments
This work was supported by Sidney Sussex College Cambridge (G.M.), the Swiss National Science Foundation (T.C.T.M.), Peterhouse College, Cambridge (T.C.T.M.) the ERC (T.P.J.K., S.L.), Swedish research council (S.L.), the BBSRC (T.P.J.K.), and the Newman Foundation (T.P.J.K.). We thank the members of the Knowles and Linse research groups for their input on experiences with this method, in particular Alexander Dear, Erik Hellstrand, Risto Cukalevski, Xiaoting Yang, Kalyani Sanagavarapu, Tanja Weiffert, Celine Galvagnion, Ricardo Gaspar, Ryan Limbocker, Catherine Xu and Patrick Flagmeier.
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Meisl, G., Michaels, T.C.T., Linse, S., Knowles, T.P.J. (2018). Kinetic Analysis of Amyloid Formation. In: Sigurdsson, E., Calero, M., Gasset, M. (eds) Amyloid Proteins. Methods in Molecular Biology, vol 1779. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7816-8_12
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DOI: https://doi.org/10.1007/978-1-4939-7816-8_12
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