{"data":{"id":"10.6084/m9.figshare.1245030.v3","type":"dois","attributes":{"doi":"10.6084/m9.figshare.1245030.v3","prefix":"10.6084","suffix":"m9.figshare.1245030.v3","identifiers":[],"alternateIdentifiers":[],"creators":[{"name":"Müller, Henrik","nameType":"Personal","givenName":"Henrik","familyName":"Müller","affiliation":[],"nameIdentifiers":[]},{"name":"Brener, Oleksandr","nameType":"Personal","givenName":"Oleksandr","familyName":"Brener","affiliation":[],"nameIdentifiers":[]},{"name":"Andreoletti, Olivier","nameType":"Personal","givenName":"Olivier","familyName":"Andreoletti","affiliation":[],"nameIdentifiers":[]},{"name":"Piechatzek, Timo","nameType":"Personal","givenName":"Timo","familyName":"Piechatzek","affiliation":[],"nameIdentifiers":[]},{"name":"Willbold, Dieter","nameType":"Personal","givenName":"Dieter","familyName":"Willbold","affiliation":[],"nameIdentifiers":[]},{"name":"Legname, Giuseppe","nameType":"Personal","givenName":"Giuseppe","familyName":"Legname","affiliation":[],"nameIdentifiers":[]},{"name":"Heise, Henrike","nameType":"Personal","givenName":"Henrike","familyName":"Heise","affiliation":[],"nameIdentifiers":[]}],"titles":[{"title":"Progress towards structural understanding of infectious sheep PrP-amyloid"}],"publisher":"Taylor \u0026 Francis","container":{},"publicationYear":2015,"subjects":[{"subject":"Computational Biology"},{"subject":"110309 Infectious Diseases","subjectScheme":"FOR"},{"subject":"FOS: Health sciences","schemeUri":"http://www.oecd.org/science/inno/38235147.pdf","subjectScheme":"Fields of Science and Technology (FOS)"},{"subject":"FOS: Health sciences","subjectScheme":"Fields of Science and Technology (FOS)"},{"subject":"Biological Sciences"},{"subject":"Chemistry"},{"subject":"Neuroscience"},{"subject":"Biochemistry"},{"subject":"Biophysics"}],"contributors":[],"dates":[{"date":"2015-10-08","dateType":"Created"},{"date":"2016-01-19","dateType":"Updated"},{"date":"2015","dateType":"Issued"}],"language":null,"types":{"ris":"DATA","bibtex":"misc","citeproc":"dataset","schemaOrg":"Dataset","resourceType":"Dataset","resourceTypeGeneral":"Dataset"},"relatedIdentifiers":[{"relationType":"IsSupplementTo","relatedIdentifier":"10.4161/19336896.2014.983754","relatedIdentifierType":"DOI"},{"relationType":"IsIdenticalTo","relatedIdentifier":"10.6084/m9.figshare.1245030","relatedIdentifierType":"DOI"}],"relatedItems":[],"sizes":["6510612 Bytes"],"formats":[],"version":null,"rightsList":[{"rights":"Creative Commons Attribution 4.0 International","rightsUri":"https://creativecommons.org/licenses/by/4.0/legalcode","schemeUri":"https://spdx.org/licenses/","rightsIdentifier":"cc-by-4.0","rightsIdentifierScheme":"SPDX"}],"descriptions":[{"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.","descriptionType":"Abstract"}],"geoLocations":[],"fundingReferences":[],"xml":"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