Low density subcellular fractions enhance disease-specific prion protein misfolding

Graham, James F. and Agarwal, Sonya and Kurian, Dominic and Kirby, Louise and Pinheiro, Teresa J. T. and Gill, Andrew C. (2010) Low density subcellular fractions enhance disease-specific prion protein misfolding. Journal of Biological Chemistry, 285 (13). pp. 9868-9880. ISSN 1083-351X

Full content URL: http://doi.org/10.1074/jbc.M109.093484

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Abstract

The production of prion particles in vitro by amplification with or without exogenous seed typically results in infectivity titers less than those associated with PrPSc isolated ex vivo and highlights the potential role of co-factors that can catalyze disease-specific prion protein misfolding in vivo. We used a cell-free conversion assay previously shown to replicate many aspects of transmissible spongiform encephalopathy disease to investigate the cellular location of disease-specific co-factors using fractions derived from gradient centrifugation of a scrapie-susceptible cell line. Fractions from the low density region of the gradient doubled the efficiency of conversion of recombinant PrP. These fractions contain plasma membrane and cytoplasmic proteins, and conversion enhancement can be achieved using PrPSc derived from two different strains of mouse-passaged scrapie as seed. Equivalent fractions from a second scrapie-susceptible cell line also stimulate conversion. We also show that subcellular fractions enhancing disease-specific prion protein conversion prevent in vitro fibrillization of recombinant prion protein, suggesting the existence of separate, competing mechanisms of disease-specific and nonspecific misfolding in vivo.

Keywords:Neurodegeneration, prion, protein misfolding, Protein Structure, Secondary, subcellular fractionation, cell free conversion, fibrillisation
Subjects:B Subjects allied to Medicine > B140 Neuroscience
C Biological Sciences > C760 Biomolecular Science
D Veterinary Sciences, Agriculture and related subjects > D320 Animal Health
C Biological Sciences > C770 Biophysical Science
Divisions:College of Science
ID Code:29480
Deposited On:16 Mar 2018 16:57

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