Abstract
Structural investigation of the species present during protein fibrillation is of tremendous importance, yet complicated by the equilibrium between species of very different sizes and life-times. Small-angle X-ray scattering may be applied to solve this problem, providing both information about the process (number of species present and volume fractions of individual species) and low-resolution three-dimensional shape reconstructions of individual species. Here, we describe in detail the challenges associated with the approach, exemplified using data from fibrillating insulin or α-synuclein samples.
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Acknowledgments
The experiences gathered in this chapter were not collected overnight, thus we would like to thank our colleagues and collaborators for significant discussions and help. In particular, from the University of Copenhagen: Minna Grønning, Marco van de Weert, and Vito Foderá. From EMBL, Hamburg: Dmitri I. Svergun, Manfred Roessle and the rest of BioSAXS group for valuable help with programs and for being very helpful beamline contacts (which, as stated many times above, is of great importance during this kind of experiments). Likewise, we are grateful for interest and kind support from Novo Nordisk A/S. We greatly acknowledge plenty beamtime at beamline X33 (EMBL-Hamburg/DESY), without which the development of the methodology described here would not have been possible. We also appreciate funding from The Lundbeck Foundation, the Danish Council for Independent Research | Medical Sciences, and DanScatt.
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Langkilde, A.E., Vestergaard, B. (2012). Structural Characterization of Prefibrillar Intermediates and Amyloid Fibrils by Small-Angle X-Ray Scattering. In: Sigurdsson, E., Calero, M., Gasset, M. (eds) Amyloid Proteins. Methods in Molecular Biology, vol 849. Humana Press. https://doi.org/10.1007/978-1-61779-551-0_10
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DOI: https://doi.org/10.1007/978-1-61779-551-0_10
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