α-Synuclein Fibrillogenesis Is Nucleation-dependent

Wood, Stephen; Wypych, Jette; Steavenson, Shirley; Louis, Jean‐Claude; Citron, Martin; Biere, Anja Leona

doi:10.1074/jbc.274.28.19509

Cited by 646 publications

(312 citation statements)

References 28 publications

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“…4A). Under the experimental conditions we reproduced the previously reported loss of soluble ␣-synuclein, which is accompanied by the formation of fibrils (25); the depletion of soluble monomer continues until the critical concentration is reached (26). In contrast, we did not observe a loss of soluble material for either ␤-or ␥-synuclein at the same concentration (Fig.…”

Section: Resultssupporting

confidence: 73%

“…In our previously established in vitro system ␣-synuclein forms fibrillar aggregates during extended incubations by a nucleation-dependent mechanism, and both PD mutations accelerate this aggregation (to a different extent) (25,26). We now followed a similar aggregation time course to compare ␣-, ␤-, and ␥-synuclein in parallel at the same concentrations (Fig.…”

Section: Resultsmentioning

confidence: 98%

“…In vitro studies have shown that recombinant ␣-synuclein can form Lewy body-like fibrils (21-25) by a nucleation-dependent mechanism (26), and oligomeric intermediates have been described in vitro (21,27,28). Most importantly, PD-linked ␣-synuclein mutations accelerate this aggregation process (24,25), which suggests that such in vitro studies can have relevance for explaining aspects of PD pathogenesis.…”

Section: Parkinson's Disease (Pd)mentioning

confidence: 99%

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Parkinson's Disease-associated α-Synuclein Is More Fibrillogenic than β- and γ-Synuclein and Cannot Cross-seed Its Homologs

Biere

Wood

Wypych

et al. 2000

Journal of Biological Chemistry

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168

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Parkinson's disease (PD) is a neurodegenerative disorder that is pathologically characterized by the presence of intracytoplasmic Lewy bodies. Recently, two point mutations in ␣-synuclein were found to be associated with familial PD, but as of yet no mutations have been described in the homologous genes ␤-and ␥-synuclein. ␣-Synuclein forms the major fibrillar component of Lewy bodies, but these do not stain for ␤-or ␥-synuclein. This result is very surprising, given the extent of sequence conservation and the high similarity in expression and subcellular localization, in particular between ␣-and ␤-synuclein. Here we compare in vitro fibrillogenesis of all three purified synucleins. We show that fresh solutions of ␣-, ␤-, and ␥-synuclein show the same natively unfolded structure. While over time ␣-synuclein forms the previously described fibrils, no fibrils could be detected for ␤-and ␥-synuclein under the same conditions. Most importantly, ␤-and ␥-synuclein could not be cross-seeded with ␣-synuclein fibrils. However, under conditions that drastically accelerate aggregation, ␥-synuclein can form fibrils with a lag phase roughly three times longer than ␣-synuclein. These results indicate that ␤-and ␥-synuclein are intrinsically less fibrillogenic than ␣-synuclein and cannot form mixed fibrils with ␣-synuclein, which may explain why they do not appear in the pathological hallmarks of PD, although they are closely related to ␣-synuclein and are also abundant in brain. Parkinson's disease (PD)1 is a neurodegenerative disorder that predominantly affects dopaminergic neurons in the nigrostriatal system but also affects several other regions of the brain. Pathological hallmarks of PD are Lewy bodies and Lewy neurites (1-3), which also accumulate in dementia with Lewy bodies (4) but not in a variety of other neurodegenerative disorders. Recently, two dominant mutations in ␣-synuclein have been linked to familial early onset PD (5, 6). This has put ␣-synuclein at the center of investigations into the pathogenesis of PD.␣-Synuclein is closely related to two other proteins, ␤-and ␥-synuclein (Fig. 1A). With 78% similarity ␤-synuclein has been called an "almost carbon copy" of ␣-synuclein (7), and it was not trivial to generate antibodies that clearly distinguish both forms (8); ␥-synuclein shares 60% similarity at the amino acid level with ␣-synuclein (Fig. 1A). All three synucleins are highly expressed in the human brain and show a strikingly similar regional distribution. They are all expressed in the thalamus, substantia nigra, caudate nucleus, amygdala, and the hippocampus (9). Moreover, ␣-and ␤-synuclein even share the same subcellular distribution; they colocalize to presynaptic terminals in primary hippocampal neurons (10), and they show a virtually complete overlap in human and mouse brain sections as demonstrated by double-stained confocal microscopy (11). No ␣-or ␤-synuclein-specific synapses were identified (11). The high expression of ␤-and ␥-synuclein in the substantia nigra and their similarity to ␣-synuclein ...

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Section: Resultssupporting

confidence: 73%

Section: Resultsmentioning

confidence: 98%

Section: Parkinson's Disease (Pd)mentioning

confidence: 99%

See 1 more Smart Citation

Parkinson's Disease-associated α-Synuclein Is More Fibrillogenic than β- and γ-Synuclein and Cannot Cross-seed Its Homologs

Biere

Wood

Wypych

et al. 2000

Journal of Biological Chemistry

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168

143

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“…9, where increasing the number of seeds results in a shorter lag time. 26,29,68,69 Our simulations show vivid details of molecular mechanism behind the seeding phenomenon in addition to capturing the essential aspects of experimental observations.…”

Section: Seedingmentioning

confidence: 99%

Simulations of nucleation and elongation of amyloid fibrils

Zhang

Muthukumar

2009

The Journal of Chemical Physics

121

109

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We present a coarse-grained model for the growth kinetics of amyloid fibrils from solutions of peptides and address the fundamental mechanism of nucleation and elongation by using a lattice Monte Carlo procedure. We reproduce the three main characteristics of nucleation of amyloid fibrils: ͑1͒ existence of lag time, ͑2͒ occurrence of a critical concentration, and ͑3͒ seeding. We find the nucleation of amyloid fibrils to require a quasi-two-dimensional configuration, where a second layer of ␤ sheet must be formed adjunct to a first layer, which in turn leads to a highly cooperative nucleation barrier. The elongation stage is found to involve the Ostwald ripening ͑evaporation-condensation͒ mechanism, whereby bigger fibrils grow at the expense of smaller ones. This new mechanism reconciles the debate as to whether protofibrils are precursors or monomer reservoirs. We have systematically investigated the roles of time, peptide concentration, temperature, and seed size. In general, we find that there are two kinds of lag time arising from two different mechanisms. For higher temperatures or low enough concentrations close to the disassembly boundary, the fibrillization follows the nucleation mechanism. However, for low temperatures, where the nucleation time is sufficiently short, there still exists an apparent lag time due to slow Ostwald ripening mechanism. Consequently, the lag time is nonmonotonic with temperature, with the shortest lag time occurring at intermediate temperatures, which in turn depend on the peptide concentration. While the nucleation dominated regime can be controlled by seeding, the Ostwald ripening regime is insensitive to seeding. Simulation results from our coarse-grained model on the fibril size, lag time, elongation rate, and solubility are consistent with available experimental observations on many specific amyloid systems.

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“…Single point mutations in ␣-synuclein have been linked to rare familial forms of PD (28,29), but the vast majority of Lewy body-related disease cases apparently involve the wildtype form of ␣-synuclein (30 -36). Purified synucleins will aggregate in vitro into fibrils resembling those found in Lewy bodies, following an initial nucleation step in which smaller synuclein multimers are formed (37)(38)(39)(40)(41). In healthy brain tissue, multimeric forms of synuclein are not typically observed, but multimers are prominent in brain tissue from individuals afflicted with Lewy body diseases (15,17,42,43).…”

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confidence: 99%