Abstract
A fluorescence technology to monitor the proliferation of amyloidogenic neurological disorders is proposed. A crude brain homogenate (0.01%) from animals infected with a transmissible spongiform encephalopathy is employed as a catalytic medium initiating conformational changes in 520 nM polypeptide biosensors (Tris/trifluoroethanol 50% mixture at pH 7). The fluorescence methods utilize pyrene residues covalently attached to the peptide ends. The coil-to-β-strand transitions in biosensor molecules cause elevation of a distinct fluorescence band of the pyrene aggregates (i.e. excimers). This approach enables the detection of infectious prion proteins at fmol, does not require antibody binding or protease treatment. Technology might be adopted for diagnosing a large variety of conformational disorders as well as for generic high-throughput screening of the amyloidogenic potential in plasma.
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This work was supported by the NIH grant (1R43 HL70399-01).
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Tcherkasskaya, O., Davidson, E.A., Schmerr, M.J. et al. Conformational biosensor for diagnosis of prion diseases. Biotechnol Lett 27, 671–675 (2005). https://doi.org/10.1007/s10529-005-4478-7
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DOI: https://doi.org/10.1007/s10529-005-4478-7