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Free keywords:
Parkinson's disease; magic-angle spinning; sequential assignment
strategy; secondary structure; supramolecular arrangement
Abstract:
Fibrillar
α
-synuclein (AS) is the major component of Lewy bodies, the
pathological hallmark of Parkinson's disease. Mouse AS (mAS) aggregates
much faster than human AS (hAS), although mAS differs from hAS at only
seven positions in its primary sequence. Currently, little is known about the
site-speci
fi
c structural differences between mAS and hAS
fi
brils. Here, we
applied state-of-the-art solid-state nuclear magnetic resonance (ssNMR)
methods to structurally characterize mAS
fi
brils. The assignment strategy
employed a set of high-resolution 2D and 3D ssNMR spectra recorded on
uniformly [
13
C,
15
N], [1-
13
C]glucose, and [2-
13
C]glucose labeled mAS
fi
brils.
An almost complete resonance assignment (96% of backbone amide
15
Nand
93% of all
13
C nuclei) was obtained for residues from Gly41 to Val95, which
form the core of mAS
fi
brils. Six
β
-strands were identi
fi
ed to be within the
fi
bril core of mAS based on a secondary chemical shift and NHHC analysis.
Intermolecular
13
C:
15
N labeled restraints obtained from mixed 1:1
13
C/
15
N-
labeled mAS
fi
brils reveal a parallel, in-register supramolecular
β
-sheet
arrangement. The results were compared in detail to recent structural studies
on hAS
fi
brils and indicate the presence of a structurally conserved motif
comprising residues Glu61–Lys80.
