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Progress towards structural understanding of infectious sheep PrP-amyloid
The still elusive structural difference of non-infectious and infectious amyloid of the mammalian prion protein (PrP) is a major pending milestone in understanding protein-mediated infectivity in neurodegenerative diseases. Preparations of PrP-amyloid proven to be infectious have never been investig...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4601355/ https://www.ncbi.nlm.nih.gov/pubmed/25482596 http://dx.doi.org/10.4161/19336896.2014.983754 |
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author | Müller, Henrik Brener, Oleksandr Andreoletti, Olivier Piechatzek, Timo Willbold, Dieter Legname, Giuseppe Heise, Henrike |
author_facet | Müller, Henrik Brener, Oleksandr Andreoletti, Olivier Piechatzek, Timo Willbold, Dieter Legname, Giuseppe Heise, Henrike |
author_sort | Müller, Henrik |
collection | PubMed |
description | The still elusive structural difference of non-infectious and infectious amyloid of the mammalian prion protein (PrP) is a major pending milestone in understanding protein-mediated infectivity in neurodegenerative diseases. Preparations of PrP-amyloid proven to be infectious have never been investigated with a high-resolution technique. All available models to date have been based on low-resolution data. Here, we establish protocols for the preparation of infectious samples of full-length recombinant (rec) PrP-amyloid in NMR-sufficient amounts by spontaneous fibrillation and seeded fibril growth from brain extract. We link biological and structural data of infectious recPrP-amyloid, derived from bioassays, atomic force microscopy, and solid-state NMR spectroscopy. Our data indicate a semi-mobile N‑terminus, some residues with secondary chemical shifts typical of α‑helical secondary structure in the middle part between ∼115 to ∼155, and a distinct β‑sheet core C‑terminal of residue ∼155. These findings are not in agreement with all current models for PrP-amyloid. We also provide evidence that samples seeded from brain extract may not differ in the overall arrangement of secondary structure elements, but rather in the flexibility of protein segments outside the β-core region. Taken together, our protocols provide an essential basis for the high-resolution characterization of non-infectious and infectious PrP-amyloid in the near future. |
format | Online Article Text |
id | pubmed-4601355 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-46013552015-11-18 Progress towards structural understanding of infectious sheep PrP-amyloid Müller, Henrik Brener, Oleksandr Andreoletti, Olivier Piechatzek, Timo Willbold, Dieter Legname, Giuseppe Heise, Henrike Prion Research Paper The still elusive structural difference of non-infectious and infectious amyloid of the mammalian prion protein (PrP) is a major pending milestone in understanding protein-mediated infectivity in neurodegenerative diseases. Preparations of PrP-amyloid proven to be infectious have never been investigated with a high-resolution technique. All available models to date have been based on low-resolution data. Here, we establish protocols for the preparation of infectious samples of full-length recombinant (rec) PrP-amyloid in NMR-sufficient amounts by spontaneous fibrillation and seeded fibril growth from brain extract. We link biological and structural data of infectious recPrP-amyloid, derived from bioassays, atomic force microscopy, and solid-state NMR spectroscopy. Our data indicate a semi-mobile N‑terminus, some residues with secondary chemical shifts typical of α‑helical secondary structure in the middle part between ∼115 to ∼155, and a distinct β‑sheet core C‑terminal of residue ∼155. These findings are not in agreement with all current models for PrP-amyloid. We also provide evidence that samples seeded from brain extract may not differ in the overall arrangement of secondary structure elements, but rather in the flexibility of protein segments outside the β-core region. Taken together, our protocols provide an essential basis for the high-resolution characterization of non-infectious and infectious PrP-amyloid in the near future. Taylor & Francis 2014-11-18 /pmc/articles/PMC4601355/ /pubmed/25482596 http://dx.doi.org/10.4161/19336896.2014.983754 Text en © 2014 The Author(s). Published with license by Taylor & Francis http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted. |
spellingShingle | Research Paper Müller, Henrik Brener, Oleksandr Andreoletti, Olivier Piechatzek, Timo Willbold, Dieter Legname, Giuseppe Heise, Henrike Progress towards structural understanding of infectious sheep PrP-amyloid |
title | Progress towards structural understanding of infectious sheep PrP-amyloid |
title_full | Progress towards structural understanding of infectious sheep PrP-amyloid |
title_fullStr | Progress towards structural understanding of infectious sheep PrP-amyloid |
title_full_unstemmed | Progress towards structural understanding of infectious sheep PrP-amyloid |
title_short | Progress towards structural understanding of infectious sheep PrP-amyloid |
title_sort | progress towards structural understanding of infectious sheep prp-amyloid |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4601355/ https://www.ncbi.nlm.nih.gov/pubmed/25482596 http://dx.doi.org/10.4161/19336896.2014.983754 |
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