TY  - JOUR
AU  - Müller, Henrik
AU  - Brener, Oleksandr
AU  - Andreoletti, Olivier
AU  - Piechatzek, Timo
AU  - Willbold, Dieter
AU  - Legname, Giuseppe
AU  - Heise, Henrike
TI  - Progress towards structural understanding of infectious sheep PrP-amyloid
JO  - Prion
VL  - 8
IS  - 5
SN  - 1933-690X
CY  - Austin, Tex.
PB  - Landes Bioscience
M1  - FZJ-2014-06732
SP  - 344-358
PY  - 2014
AB  - 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.
LB  - PUB:(DE-HGF)16
UR  - <Go to ISI:>//WOS:000348376700004
DO  - DOI:10.4161/19336896.2014.983754
UR  - https://juser.fz-juelich.de/record/173322
ER  -