Abstract
Oligomeric assemblies of the amyloid β-protein, Aβ, are thought to be the proximate neurotoxic agents in Alzheimer’s disease (AD). Oligomer formation is a complex process that produces a polydisperse population of metastable structures. For this reason, formal structure–activity correlations, both in vitro and in vivo, have been difficult to accomplish. An analytical solution to this problem was provided by the application of a photochemical cross-linking method to the Aβ assembly system. This method, photo-induced cross-linking of unmodified proteins (PICUP), enabled the quantitative determination of the oligomer size distribution. We report here the integration of PICUP with SDS-PAGE and alkaline extraction procedures to create a method for the isolation of pure populations of oligomers of defined order. This method has been used successfully to provide material for formal structure–activity studies of Aβ oligomers.
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Acknowledgments
We gratefully acknowledge the support of NIH grants AG027818, NS038328, the Jim Easton Consortium for Alzheimer’s Drug Discovery and Biomarkers at UCLA, and the California Department of Public Health, Alzheimer’s Disease Program, grant #07-65806.
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Rosensweig, C., Ono, K., Murakami, K., Lowenstein, D.K., Bitan, G., Teplow, D.B. (2012). Preparation of Stable Amyloid β-Protein Oligomers of Defined Assembly Order. 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_3
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DOI: https://doi.org/10.1007/978-1-61779-551-0_3
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