Formation of toxic oligomeric alpha-synuclein species in living cells. PLoS One 2008 Apr 02;3(4):e1867
Date
04/03/2008Pubmed ID
18382657Pubmed Central ID
PMC2270899DOI
10.1371/journal.pone.0001867Scopus ID
2-s2.0-44849125207 (requires institutional sign-in at Scopus site) 386 CitationsAbstract
BACKGROUND: Misfolding, oligomerization, and fibrillization of alpha-synuclein are thought to be central events in the onset and progression of Parkinson's disease (PD) and related disorders. Although fibrillar alpha-synuclein is a major component of Lewy bodies (LBs), recent data implicate prefibrillar, oligomeric intermediates as the toxic species. However, to date, oligomeric species have not been identified in living cells.
METHODOLOGY/PRINCIPAL FINDINGS: Here we used bimolecular fluorescence complementation (BiFC) to directly visualize alpha-synuclein oligomerization in living cells, allowing us to study the initial events leading to alpha-synuclein oligomerization, the precursor to aggregate formation. This novel assay provides us with a tool with which to investigate how manipulations affecting alpha-synuclein aggregation affect the process over time. Stabilization of alpha-synuclein oligomers via BiFC results in increased cytotoxicity, which can be rescued by Hsp70 in a process that reduces the formation of alpha-synuclein oligomers. Introduction of PD-associated mutations in alpha-synuclein did not affect oligomer formation but the biochemical properties of the mutant alpha-synuclein oligomers differ from those of wild type alpha-synuclein.
CONCLUSIONS/SIGNIFICANCE: This novel application of the BiFC assay to the study of the molecular basis of neurodegenerative disorders enabled the direct visualization of alpha-synuclein oligomeric species in living cells and its modulation by Hsp70, constituting a novel important tool in the search for therapeutics for synucleinopathies.
Author List
Outeiro TF, Putcha P, Tetzlaff JE, Spoelgen R, Koker M, Carvalho F, Hyman BT, McLean PJMESH terms used to index this publication - Major topics in bold
AnimalsCHO Cells
Cell Line, Tumor
Cricetinae
Cricetulus
HSP70 Heat-Shock Proteins
Humans
Lewy Bodies
Microscopy, Fluorescence
Mutation
Parkinson Disease
Protein Folding
Protein Structure, Tertiary
alpha-Synuclein
