Effects of Variable Oligosaccharide Lengths on the Infectious Nature of Chronic Wasting Disease Prion Proteins

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Authors

Breeggemann, Matthew

Date of Issue

2012-04-01

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thesis

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Prion, disease, oligosaccharides

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Abstract

The rate at which a disease causing prion protein (PrPSc) converts a normal prion protein (PrPC) to the infectious form may depend on the presence of terminal sugar moieties on the oligosaccharides of the infectious or normal protein. The first goal of this project was to use two different insect cell lines in order to evaluate and compare the ability of these cells to produce PrPC. The second goal was to evaluate the stability of prion protein from these cells one year after expression when stored at -20°C. The next objective was to assess if the proteins are GPI anchored and exploit this feature to extract them from cell membranes. The fourth goal was to use lectin affinity interactions to evaluate the differences in glycosylation status between the prion proteins produced from these two cell lines. Ultimately, protein misfolding conversion assays (PMCA) will be used to evaluate any differences in conversion rates between the fully glycosylated and partially glycosylated prions. The results of these experiments may provide insight into the mechanism for conversion of normal prions into disease causing ones by elucidating the role of oligosaccharides in the conversion process.

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