Biasing the native α-synuclein conformational ensemble towards compact states abolishes aggregation and neurotoxicity

The aggregation of α-synuclein (α-syn) into amyloid fibrils is a major pathological hallmark of Parkinson's disease (PD) and other synucleinopathies. The mechanisms underlying the structural transition of soluble and innocuous α-syn to aggregated neurotoxic forms remains largely unknown. The di...

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Bibliographic Details
Authors: Carija, Anita|||0000-0001-5972-7448, Garcia de Carvalho Pinheiro, Francisca|||0000-0003-3778-1528, Pujols Pujol, Jordi|||0000-0001-9424-5866, Brás, Inês C., Lázaro, Diana Fernandes, Santambrogio, Carlo, Grandori, Rita, Outeiro, Tiago Fleming, Navarro, Susanna|||0000-0001-8160-9536, Ventura, Salvador|||0000-0002-9652-6351
Format: article
Publication Date:2019
Country:España
Institution:Universitat Autònoma de Barcelona
Repository:Dipòsit Digital de Documents de la UAB
Language:English
OAI Identifier:oai:ddd.uab.cat:226349
Online Access:https://ddd.uab.cat/record/226349
https://dx.doi.org/urn:doi:10.1016/j.redox.2019.101135
Access Level:Open access
Keyword:α-syn, α-synuclein
PD, Parkinson's disease
NAC, non-amyloid β-component
α-synCC, disulfide bridge-containing variant
CD, circular dichroism
Th-T, Thioflavin-T
CR, Congo Red
TEM, Transmission electron microscopy
PK, proteinase K
DOPC, dioleoylphosphatidylcholine
DMPS, dimyristoylphosphatidylserine
α-synuclein
Disulfide bond
Amyloid
Protein aggregation
Parkinson's disease
Description
Summary:The aggregation of α-synuclein (α-syn) into amyloid fibrils is a major pathological hallmark of Parkinson's disease (PD) and other synucleinopathies. The mechanisms underlying the structural transition of soluble and innocuous α-syn to aggregated neurotoxic forms remains largely unknown. The disordered nature of α-syn has hampered the use of structure-based protein engineering approaches to elucidate the molecular determinants of this transition. The recent 3D structure of a pathogenic α-syn fibril provides a template for this kind of studies. The structure supports the NAC domain being a critical element in fibril formation, since it constitutes the core of the fibril, delineating a Greek-key motif. Here, we stapled the ends of this motif with a designed disulfide bond and evaluated its impact on the conformation, aggregation and toxicity of α-syn in different environments. The new covalent link biases the native structural ensemble of α-syn toward compact conformations, reducing the population of fully unfolded species. This conformational bias results in a strongly reduced fibril formation propensity both in the absence and in the presence of lipids and impedes the formation of neurotoxic oligomers. Our study does not support the Greek-key motif being already imprinted in early α-syn assemblies, discarding it as a druggable interface to prevent the initiation of fibrillation. In contrast, it suggests the stabilization of native, compact ensembles as a potential therapeutic strategy to avoid the formation of toxic species and to target the early stages of PD.