The Study of Amyloid Fibril Formation of Full-length Mouse Prion Protein and Short Mouse Prion Peptide
| Autor: | Chung-Yu Lee, 李忠諭 |
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| Rok vydání: | 2009 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 97 Amyloids are aggregates of misfolded proteins. Many diseases are caused by amyloids accumulation. The prion disease, also called transmissible spongiform encephalopathy (TSE), is caused by conformational change of host-encoded prion protein to a β-sheet-rich conformer. When prion protein changes its conformation, it will aggregate and form fibril-like structure. To know how prion protein forms amyloid fibrils and the factors affecting its formation, we expressed full-length mouse prion protein [mPrP(23-231)] in Escherichia coli and purified it. We studied the amyloid fibril formation in vitro by thioflavin T (ThT) fluorescence spectroscopy. In mild denaturing condition with vigorous shaking, mPrP(23-231) forms amyloid fibrils after one-day incubation. Adding pre-formed amyloid fibrils as seeds may shorten the lag time of amyloid fibril formation. In addition, we synthesized mouse prion peptide (mPrP(108-144)) to do cross seeding experiment. Full-length mPrP(23-231) can function as seeds to accelerate the fibril formation of chemically synthetic mPrP(108-144), and vice versa. Amyloid fibrils formed from mPrP(23-231) and mPrP(108-144) with share very similar morphology in TEM and fluorescence lifetime assay using ThT as dye, suggesting that mPrP(23-231) and mPrP(108-144) has the same amyloid core. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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