Abstract
Conformational conversion of proteins in disease is likely to be accompanied by molecular surface exposure of previously sequestered amino-acid side chains. We found that induction of β-sheet structures in recombinant prion proteins is associated with increased solvent accessibility of tyrosine. Antibodies directed against the prion protein repeat motif, tyrosine-tyrosine-arginine, recognize the pathological isoform of the prion protein but not the normal cellular isoform, as assessed by immunoprecipitation, plate capture immunoassay and flow cytometry. Antibody binding to the pathological epitope is saturable and specific, and can be created in vitro by partial denaturation of normal brain prion protein. Conformation-selective exposure of Tyr-Tyr-Arg provides a probe for the distribution and structure of pathologically misfolded prion protein, and may lead to new diagnostics and therapeutics for prion diseases.
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Acknowledgements
We thank A. Aguzzi, C. Bergeron, R. Jackman, R. Carp, B. Oesch, R. Rohwer, R. Rubinstein and M. J. Schmerr for provision of infected material and facilities; K. Qin and D. Westaway for recombinant mouse PrP; B. Bartol, P. Cunningham, P. Cecchetti, B. O'Brien-Graf, C. Quan and E. Thibaudeau for expert technical assistance; D. Chelsky and J. Griffin for critical reading of the manuscript; and C. Desjardins and L. Segal for their encouragement and support of this work. N.R.C. is the Jeno and Ilona Diener Chair of Neurodegenerative Diseases at the University of Toronto and Sunnybrook & Women's College Health Sciences Center, and is a Founder and Scientific Advisor of Caprion Pharmaceuticals. This work was supported by Caprion Pharmaceuticals, the Canadian Institutes of Health Research (Institute of Infection and Immunity) and McDonald's Corporation.
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The manuscript represents university-industry collaboration between scientists at the University of Toronto and two biotechnology companies (Caprion Pharmaceuticals of Montreal, Canada and Idexx Laboratories of Portland, Maine). N.R.C. is a professor at the University of Toronto, as well as founder and scientific advisor of Caprion Pharmaceuticals, from which he has derived partial salary and laboratory operating grants. The commercial aspects of this discovery are being developed under license from Caprion to Idexx and Ortho Clinical Diagnostics.
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Paramithiotis, E., Pinard, M., Lawton, T. et al. A prion protein epitope selective for the pathologically misfolded conformation. Nat Med 9, 893–899 (2003). https://doi.org/10.1038/nm883
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DOI: https://doi.org/10.1038/nm883
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