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@ARTICLE{Mller:173322,
author = {Müller, Henrik and Brener, Oleksandr and Andreoletti,
Olivier and Piechatzek, Timo and Willbold, Dieter and
Legname, Giuseppe and Heise, Henrike},
title = {{P}rogress towards structural understanding of infectious
sheep {P}r{P}-amyloid},
journal = {Prion},
volume = {8},
number = {5},
issn = {1933-690X},
address = {Austin, Tex.},
publisher = {Landes Bioscience},
reportid = {FZJ-2014-06732},
pages = {344-358},
year = {2014},
abstract = {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.},
cin = {ICS-6},
ddc = {570},
cid = {I:(DE-Juel1)ICS-6-20110106},
pnm = {452 - Structural Biology (POF2-452)},
pid = {G:(DE-HGF)POF2-452},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000348376700004},
doi = {10.4161/19336896.2014.983754},
url = {https://juser.fz-juelich.de/record/173322},
}