Abstract
The occurrence of multiple strains of prions may reflect conformational variability of PrPSc, a disease-associated, aggregated variant of the cellular prion protein, PrPC. Here we used luminescent conjugated polymers (LCPs), which emit conformation-dependent fluorescence spectra, for characterizing prion strains. LCP reactivity and emission spectra of brain sections discriminated among four immunohistochemically indistinguishable, serially mouse-passaged prion strains derived from sheep scrapie, chronic wasting disease (CWD), bovine spongiform encephalopathy (BSE), and mouse-adapted Rocky Mountain Laboratory scrapie prions. Furthermore, using LCPs we differentiated between field isolates of BSE and bovine amyloidotic spongiform encephalopathy, and identified noncongophilic deposits in prion-infected deer and sheep. We found that fibrils with distinct morphologies generated from chemically identical recombinant PrP yielded unique LCP spectra, suggesting that spectral characteristic differences resulted from distinct supramolecular PrP structures. LCPs may help to detect structural differences among discrete protein aggregates and to link protein conformational features with disease phenotypes.
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Acknowledgements
This study was supported by grants from the European Union (Tseur to A.A. and Understanding Protein Misfolding and Aggregation by NMR (UPMAN) to K.W.), the Swiss National Science Foundation, the Swiss National Competence Centers for Research on Neural Plasticity and Repair (A.A.) and on Structural Biology (K.W.), US National Institutes of Health K08-AI01802 (C.J.S.), the Foundation for Research at the University of Zürich (C.J.S.), the US National Prion Research Program (C.J.S. and A.A.), the Knut and Alice Wallenberg foundation (K.P.R.N.), the Swedish Foundation for Strategic Research (P.H.), the Wenner-Gren Foundations and the Swedish Research Council (P.H.), and the Natural Sciences and Engineering Research Council of Canada (M.L.). We thank P. Vilkman, S. Fransson and C. von Schroetter for providing technical support, P. Tittmann (Electron Microscopy Center, ETH Zurich) for providing electron microscopy services, the staff at the confocal microscopy centers at the ETH Zurich and at the University of Zurich for help with spectral collection, B. Seifert for help with statistical analysis, and C. Casalone (Istituto Zooprofilattico Sperimentale del Piemonte), F. Ehrensperger (University of Zürich), M. Miller (Colorado Division of Wildlife) and K. O'Rourke (United States Department of Agriculture) for BASE, BSE, CWD and sheep scrapie brain samples, respectively.
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K.P.R.N. is part owner of a company commercializing the LCPs and the methods described in this article.
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Sigurdson, C., Nilsson, K., Hornemann, S. et al. Prion strain discrimination using luminescent conjugated polymers. Nat Methods 4, 1023–1030 (2007). https://doi.org/10.1038/nmeth1131
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DOI: https://doi.org/10.1038/nmeth1131
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