Structured Regions of Alpha-synuclein Fibrils Include the Early Onset Parkinson's Disease Mutation Sites

Journal Article · Fri Aug 26 00:00:00 EDT 2011 · Journal of Molecular Biology, 411(4):881-895

DOI:https://doi.org/10.1016/j.jmb.2011.06.026· OSTI ID:1029042

Comellas Canal, Gemma; Lemkau, Luisel R; Nieuwkoop, Andrew J; Kloepper, Kathryn D; Ladror, Daniel T; Ebisu, Reika; Woods, Wendy S; Lipton, Andrew S; George, Julia M; Rienstra, Chad M

Alpha-Synuclein (AS) fibrils constitute the major proteinaceous component of Lewy bodies (LBs), the pathological hallmark of Parkinson’s disease (PD) and other neurodegenerative diseases. Three single point mutations in the AS gene, as well as multiplication of the wild-type (WT) AS allele, have been previously identified in families with early-onset PD. Although AS fibrils have been the subject of intense study, critical details about their structure including the precise location of the B-strands and the extent of the core, the three-dimensional structure and the effects of the mutations—remain unknown. Here, we have used magic-angle spinning solid-state NMR spectroscopy to present a detailed characterization of the full-length WT AS fibrils. With improved sample preparations, isotopic labeling patterns and NMR experiments, we have confidently assigned more than 90% of the 13C and 15N backbone and sidechain chemical shifts of the detected residues from residue 39 to 97, and quantified the conformational dynamics throughout this region. Our results demonstrate that the core of AS fibrils extends with a repeated motif and that residues 30, 46 and 53-the early-onset PD mutant sites-are located in structured regions of AS fibrils.

• Research Organization: Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 1029042
• Report Number(s): PNNL-SA-77470; JMOBAK; 30444; 26716; 44750; KP1704020; TRN: US201122%%673
• Journal Information: Journal of Molecular Biology, 411(4):881-895, Vol. 411, Issue 4; ISSN 0022-2836
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
CHEMICAL SHIFT
DISEASES
GENE MUTATIONS
MUTANTS
MUTATIONS
NERVOUS SYSTEM DISEASES
RESIDUES
SAMPLE PREPARATION
SPECTROSCOPY
Lewy bodies
magic-angle spinning
solid-state NMR
conformational dynamics
structural perturbations
SOLID-STATE NMR
ANGLE-SPINNING NMR
HUMAN PRION PROTEIN
AMYLOID FIBRILS
CROSS-POLARIZATION
CHEMICAL-SHIFT
CORE STRUCTURE
MUTANT FORMS
WILD-TYPE
Environmental Molecular Sciences Laboratory