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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...

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Autores principales: Müller, Henrik, Brener, Oleksandr, Andreoletti, Olivier, Piechatzek, Timo, Willbold, Dieter, Legname, Giuseppe, Heise, Henrike
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2014
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.
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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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