Structural characterization of α‐synuclein in an aggregation prone state

Cho, Min Kyu; Nodet, Gabrielle; Kim, Hai‐Young; Jensen, Malene Ringkjøbing; Bernadó, Pau; Fernandez, Claudio O.; Becker, Stefan; Blackledge, Martin; Zweckstetter, Markus

doi:10.1002/pro.194

Cited by 103 publications

(137 citation statements)

References 29 publications

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“…Dedmon et al showed strong interactions between the highly charged C terminus and the NAC region (33). Cho et al showed that compaction increased between the C terminus and the NAC at low pH and the C terminus became more rigid, a similar conclusion to our finding that diffusion slows (6). Recently Trexler and Rhoades measured various intramolecular distances by single molecule FRET and showed substantial collapse at low pH but the effect was largest for the C terminus (34).…”

Section: Discussionsupporting

confidence: 89%

“…In vitro, the aggregation propensity of α-synuclein is very sensitive to solution conditions such as temperature, pH, salt concentration, and solution cofactors. There has been significant work to discover the structures that form in association with lipids, structures of final aggregates, and structures that may be the aggregation precursor (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15). However, the underlying reason for why monomeric α-synuclein is so prone to aggregation is still unknown.…”

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confidence: 99%

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Aggregation of α-synuclein is kinetically controlled by intramolecular diffusion

Ahmad

Chen

Lapidus

2012

Proc. Natl. Acad. Sci. U.S.A.

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We hypothesize that the first step of aggregation of disordered proteins, such as α-synuclein, is controlled by the rate of backbone reconfiguration. When reconfiguration is fast, bimolecular association is not stable, but as reconfiguration slows, association is more stable and subsequent aggregation is faster. To investigate this hypothesis, we have measured the rate of intramolecular diffusion in α-synuclein, a protein involved in Parkinson's disease, under solvent conditions that accelerate or decelerate aggregation. Using the method of tryptophan-cysteine (Trp-Cys) quenching, the rate of intramolecular contact is measured in four different loops along the chain length. This intrinsically disordered protein is highly diffusive at low temperature at neutral pH, when aggregation is slow, and compacts and diffuses more slowly at high temperature or low pH, when aggregation is rapid. Diffusion also slows with the disease mutation A30P. This work provides unique insights into the earliest steps of α-synuclein aggregation pathway and should provide the basis for the development of drugs that can prevent aggregation at the initial stage.unfolded state | Alzheimer's disease | amyloid T he protein α-synuclein is the primary component of intracellular aggregants known as Lewy bodies, a hallmark of Parkinson's disease, and likely plays a central role in neuronal cell death (1). When recombinantly expressed and purified it is intrinsically disordered in solution and highly prone to aggregation (2). Recent work has shown that the native structure in human cells is likely a helical tetramer that is resistant to aggregation, but when this tetramer is denatured the monomers cannot autonomously refold and aggregate very quickly (3, 4). Thus it is likely that the aggregation pathway passes through the monomer. In vitro, the aggregation propensity of α-synuclein is very sensitive to solution conditions such as temperature, pH, salt concentration, and solution cofactors. There has been significant work to discover the structures that form in association with lipids, structures of final aggregates, and structures that may be the aggregation precursor (5-15). However, the underlying reason for why monomeric α-synuclein is so prone to aggregation is still unknown. We propose that a physical influence on aggregation is the internal dynamics of the disordered chain, that is, the rate of intramolecular diffusion.Over the past decade, a number of groups have measured intramolecular diffusion in a variety of sequences using measurements of intramolecular contact between two probes placed in the chain. All unstructured peptides, including fragments of α-synuclein, diffuse relatively rapidly, approximately 1-3 × 10 −6 cm 2 s −1 (16-18). These rates are less than 10 times slower than free diffusion of individual amino acids and about the same as translational diffusion of the entire chain, suggesting that the chain does not exert significant drag on the movement of individual segments (19), although there are measurable tail effects when ...

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Section: Discussionsupporting

confidence: 89%

mentioning

confidence: 99%

Aggregation of α-synuclein is kinetically controlled by intramolecular diffusion

Ahmad

Chen

Lapidus

2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Both SEC and native gels separate proteins based on molecular size and shape, and our findings illustrate a key physical feature of disordered proteins-that they are significantly more extended than globular proteins of the same MW. This has been shown to be true for disordered monomer AS by several different experimental methods, 7,8 including native gel electrophoresis. 9 Moreover, it emphasizes that caution must be used in interpreting molecular size from methods that are dependent upon the shape of the molecules measured.…”

Section: Resultsmentioning

confidence: 79%

N‐terminal acetylation is critical for forming α‐helical oligomer of α‐synuclein

2012

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The aggregation of the protein a-synuclein (AS) is critical to the pathogenesis of Parkinson's disease. Although generally described as an unstructured monomer, recent evidence suggests that the native form of AS may be an a-helical tetramer which resists aggregation. Here, we show that N-terminal acetylation in combination with a mild purification protocol results in an oligomeric form of AS with partial a-helical structure. N-terminal acetylation of AS could have important implications for both the native and pathological structures and functions of AS. Through our demonstration of a recombinant expression system, our results represent an important step toward biochemical and biophysical characterization of this potentially important form of AS.

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“…However, all evidence indicates that the aggregation pathway passes through an at least partially unstructured state. Thus, much work has focused on which subset of the conformation ensemble is prone to nucleating aggregation (3)(4)(5)(6).…”

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confidence: 99%

Molecular Basis for Preventing α-Synuclein Aggregation by a Molecular Tweezer

Acharya

Safaie

Wongkongkathep

et al. 2014

Journal of Biological Chemistry

117

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Background: The molecular tweezer, CLR01, binds to Lys and prevents aggregation of ␣-synuclein. Results: CLR01 binds directly to monomeric ␣-synuclein near the N terminus and changes the charge distribution in the sequence, swelling the chain, and increasing the protein reconfiguration rate. Conclusion: Aggregation is inhibited by making the protein more diffusive. Significance: The most effective aggregation inhibitors may change monomer dynamics rather than structure.

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Structural characterization of α‐synuclein in an aggregation prone state

Cited by 103 publications

References 29 publications

Aggregation of α-synuclein is kinetically controlled by intramolecular diffusion

Aggregation of α-synuclein is kinetically controlled by intramolecular diffusion

N‐terminal acetylation is critical for forming α‐helical oligomer of α‐synuclein

Molecular Basis for Preventing α-Synuclein Aggregation by a Molecular Tweezer

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