Journal ArticleDOI
Impact of the Acidic C-Terminal Region Comprising Amino Acids 109−140 on α-Synuclein Aggregation in Vitro†
TLDR
The effects of the C-terminus on aggregation cannot be rationalized merely by a contribution to the protein net charge, but rather suggest a specific role of aa109-140 in the regulation of aggregation, presumably involving formation of intramolecular contacts.Abstract:
The aggregation of alpha-synuclein, involved in the pathogenesis of several neurodegenerative disorders such as Parkinson's disease, is enhanced in vitro by biogenic polyamines binding to the highly charged C-terminal region aa109-140. In this study, we investigated the influence of this region on the aggregation kinetics, monitored by thioflavin T binding and static light scattering, and morphology, assessed by electron microscopy, fluorescence microscopy, and turbidity, by comparing the effect of various solution conditions on the wild-type protein, the disease related mutants A53T and A30P, and two truncated variants, syn(1-108) and syn(1-124), lacking the complete or the C-terminal half of the polyamine binding site. In the presence of the intact C-terminus, aggregation was strongly retarded in physiological buffer. This inhibition of aggregation was overridden by (i) addition of spermine or MgCl(2) or lowering of pH, leading to strong charge shielding in the C-terminus or (ii) by truncation of aa125-140 or aa109-140. Addition of MgCl(2) or spermine or acidification were not effective in promoting aggregation of syn(1-108). The impact of the disease-related mutations on the aggregation kinetics was dependent on the solution conditions, with the aggregation propensity order A53T approximately wt > A30P at low ionic strength, but A53T > wt approximately A30P at high ionic strength, with exceedingly potent promotion of aggregation by the A53T mutation in the presence of spermine. In contrast to full-length alpha-synuclein aggregates, those formed from syn(1-108) did not exhibit a pronounced polymorphism. The effects of the C-terminus on aggregation cannot be rationalized merely by a contribution to the protein net charge, but rather suggest a specific role of aa109-140 in the regulation of aggregation, presumably involving formation of intramolecular contacts.read more
Citations
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Journal ArticleDOI
Parkinson's disease as a member of Prion-like disorders.
TL;DR: The clinical and experimental data that have led to think of alpha-synuclein as a prion-like protein are described and data from in vitro, cellular and animal models supporting this view are summarized.
Journal ArticleDOI
The chaperone activity of α‐synuclein: Utilizing deletion mutants to map its interaction with target proteins
TL;DR: It is concluded that the amyloidogenic NAC region (residues 61–95) contains the chaperone‐binding site which is optimized for target protein binding as a result of its β‐sheet formation and/or ordered aggregation by α‐synuclein.
Journal ArticleDOI
The N terminus of α-synuclein dictates fibril formation.
TL;DR: In this article, the authors examined three N-terminal truncations, in which deletions of 13-, 35-, and 40 residues in the N terminus modulated both aggregation kinetics and fibril morphologies.
Journal ArticleDOI
Tri- and pentamethine cyanine dyes for fluorescent detection of α-synuclein oligomeric aggregates
Vladyslava B. Kovalska,M. Yu. Losytskyy,O. I. Tolmachev,Yu. L. Slominskii,Gezina M.J. Segers-Nolten,Vinod Subramaniam,Vinod Subramaniam,Sergiy M. Yarmoluk +7 more
TL;DR: It is proposed that wide aromatic system of SL-631 pentamethine dye molecule could better fix on the less dense and structured oligomeric formation, while less bulky and more “crescent-shape” molecule of trimethine dyed SH-299 could easier enter into the groove of beta-pleated structure.
Journal ArticleDOI
A nanobody-based fluorescent reporter reveals human α-synuclein in the cell cytosol.
Christoph Gerdes,Natalia Waal,Thomas Offner,Thomas Offner,Eugenio F. Fornasiero,Nora Wender,Hannes Verbarg,Ivan Manzini,Ivan Manzini,Claudia Trenkwalder,Brit Mollenhauer,Timo Strohaeker,Markus Zweckstetter,Markus Zweckstetter,Stefan Becker,Silvio O. Rizzoli,Fitnat Buket Basmanav,Felipe Opazo +17 more
TL;DR: A nanobody biosensor is created that reveals the presence of α-Syn in cells, which allow the detection of transmittable forms ofα-Syn present in human CSF, demonstrating the potential of FluoReSyn for clinical research and diagnostics.
References
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Journal ArticleDOI
Accelerated in vitro fibril formation by a mutant alpha-synuclein linked to early-onset Parkinson disease.
TL;DR: It is demonstrated that at higher concentrations, Lewy body-like fibrils and discrete spherical assemblies are formed; most rapidly by A53T, suggesting mutation-induced acceleration of α-synuclein fibril formation may contribute to the early onset of familial PD.
Journal ArticleDOI
Fibrils Formed in Vitro from α-Synuclein and Two Mutant Forms Linked to Parkinson's Disease are Typical Amyloid†
TL;DR: Fibrils generated in vitro from alpha-synuclein, wild-type and both mutant forms, are shown to possess very similar features that are characteristic of amyloid fibrils, including a wound and predominantly unbranched morphology, distinctive dye-binding properties, and antiparallel beta-sheet structure.
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