%0 Generic %A Müller, Henrik %A Brener, Oleksandr %A Andreoletti, Olivier %A Piechatzek, Timo %A Willbold, Dieter %A Legname, Giuseppe %A Heise, Henrike %D 2015 %T Progress towards structural understanding of infectious sheep PrP-amyloid %U https://tandf.figshare.com/articles/dataset/Progress_towards_structural_understanding_of_infectious_sheep_PrP_amyloid/1245030 %R 10.6084/m9.figshare.1245030.v3 %2 https://ndownloader.figshare.com/files/1798402 %2 https://ndownloader.figshare.com/files/1798403 %2 https://ndownloader.figshare.com/files/1798404 %2 https://ndownloader.figshare.com/files/1798405 %2 https://ndownloader.figshare.com/files/1798406 %2 https://ndownloader.figshare.com/files/1798407 %2 https://ndownloader.figshare.com/files/1798408 %2 https://ndownloader.figshare.com/files/1798409 %K middle part %K protein segments %K NMR Spectroscopy %K residue %K preparation %K core %K understanding %K Neurodegenerative diseases %K data %K structure elements %K fibril growth %K chemical shifts %K sample %K force microscopy %K prion protein %K model %X

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.

%I Taylor & Francis