The Presence of an Air–Water Interface Affects Formation and Elongation of α-Synuclein Fibrils

Campioni, Silvia; Carret, Guillaume; Jördens, Sophia; Nicoud, Lucrèce; Mezzenga, Raffaele; Riek, Roland

doi:10.1021/ja412105t

Cited by 251 publications

(249 citation statements)

References 58 publications

(145 reference statements)

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“…Expression and Purification of ␣-Synuclein-Recombinant wild-type (WT) human ␣-Syn and the ␣-Syn variant ␣-Syn(C141) were overexpressed in the Escherichia coli strain BL21 Star TM (DE3) pLysS (Invitrogen) and purified as described previously (62). 2 H, 15 N-, and 13 C, 15 N-labeled human ␣-Syn were produced using standard M9 minimal medium (63) based on D 2 O (Isotec) and H 2 O, respectively, supplemented with 3 g/liter 13 C glucose (Isotec), and 1 g/liter 15 NH 4 Cl (Isotec).…”

Section: Methodsmentioning

confidence: 99%

Preparation and Characterization of Stable α-Synuclein Lipoprotein Particles

Eichmann¹,

Campioni²,

Kowal

et al. 2016

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

Preparation and Characterization of Stable α-Synuclein Lipoprotein Particles

Eichmann¹,

Campioni²,

Kowal

et al. 2016

Journal of Biological Chemistry

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“…α-Synuclein Samples The proteins (wild type α-synuclein and the E57K variant) were expressed, purified, and prepared as samples for the experiments as described previously (17 ). The purity of the proteins was at least 98%.…”

Section: Methodsmentioning

confidence: 99%

α-Synuclein Insertion into Supported Lipid Bilayers As Seen by in Situ X-ray Reflectivity

Hähl

Möller

Kiesel

et al. 2015

ACS Chem. Neurosci.

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Large aggregates of misfolded -synuclein inside neuronal cells are the hallmarks of Parkinson's disease. The protein's natural function and its supposed toxicity, however, are believed to be closely related to its interaction with cell and vesicle membranes. Upon this interaction, the protein folds into an -helical structure and intercalates into the membrane. In this study, we focus on the changes in the lipid bilayer caused by this intrusion. In situ X-ray reflectivity was applied to determine the vertical density structure of the bilayer before and after exposure to -synuclein. It was found that the -synuclein insertion, wild type and E57K variant, caused a reduction in bilayer thickness. This effect may be one factor in the membrane pore formation ability of -synuclein. AbstractLarge aggregates of misfolded α-synuclein inside neuronal cells are the hallmarks of Parkinson's disease. The protein's natural function and its supposed toxicity, however, are believed to be closely related to its interaction with cell and vesicle membranes. Upon this interaction, the protein folds into an α-helical structure and intercalates into the membrane. In this study, we focus on the changes in the lipid bilayer caused by this intrusion. In situ X-ray reflectivity was applied to determine the vertical density structure of the bilayer before and after exposure to α-synuclein. It was found that the α-synuclein insertion, wild type and E57K variant, caused a reduction in bilayer thickness. This effect may be one factor in the membrane pore formation ability of α-synuclein.

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“…Previous work has shown that the process of nucleation of α-synuclein amyloid fibrils is likely to be heterogeneous and catalyzed by environmental features, such as air-water interfaces (26,27), lipid bilayers (28), SDS micelles and other anionic surfactants (11,29,30), or artificial interfaces such as the coatings of containers or stir bars (31). In addition, mechanical action, such as shaking and stirring, is often used to accelerate the aggregation of α-synuclein (32).…”

Section: Primary Nucleation and Fragmentation Can Be Selectively Enhamentioning

confidence: 99%

Solution conditions determine the relative importance of nucleation and growth processes in α-synuclein aggregation

Buell

Galvagnion

Gaspar³

et al. 2014

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

586

867

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The formation of amyloid fibrils by the intrinsically disordered protein α-synuclein is a hallmark of Parkinson disease. To characterize the microscopic steps in the mechanism of aggregation of this protein we have used in vitro aggregation assays in the presence of preformed seed fibrils to determine the molecular rate constant of fibril elongation under a range of different conditions. We show that α-synuclein amyloid fibrils grow by monomer and not oligomer addition and are subject to higher-order assembly processes that decrease their capacity to grow. We also find that at neutral pH under quiescent conditions homogeneous primary nucleation and secondary processes, such as fragmentation and surface-assisted nucleation, which can lead to proliferation of the total number of aggregates, are undetectable. At pH values below 6, however, the rate of secondary nucleation increases dramatically, leading to a completely different balance between the nucleation and growth of aggregates. Thus, at mildly acidic pH values, such as those, for example, that are present in some intracellular locations, including endosomes and lysosomes, multiplication of aggregates is much faster than at normal physiological pH values, largely as a consequence of much more rapid secondary nucleation. These findings provide new insights into possible mechanisms of α-synuclein aggregation and aggregate spreading in the context of Parkinson disease.seeding | prion-like behavior | neurodegenerative disease | kinetic analysis | electrostatic interactions T he conversion of soluble peptide and protein molecules into insoluble amyloid fibrils is of great interest in fields of science ranging from molecular medicine to nanotechnology (1). The formation of amyloid fibrils is a characteristic feature of a substantial number of increasingly common medical disorders, including neurodegenerative conditions such as Alzheimer's and Parkinson diseases (2). Elucidating the fundamental mechanistic steps involved in the conversion from the soluble to the fibrillar forms of the peptides and proteins involved in such disorders is crucial for understanding their origin and proliferation, and hence for exploring in a rational manner new and effective therapeutic strategies through which to combat their onset or progression (3).One general aspect of amyloid diseases is that once the first aggregates are formed it is very difficult to stop or reverse the aggregation process. This implies that aggregation needs to be studied in both the absence and presence of preformed aggregates, commonly known as seeds, to deepen our understanding of the mechanism of the self-assembly process in vivo. It is possible to define from in vitro studies the rate constants for the multiplicity of microscopic steps that increase the number and total mass of the different types of aggregates that are populated during this process. Such an analysis has recently been carried out for the Aβ42 peptide (4) associated with Alzheimer's disease by combining experimental and theoretical methodolo...

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The Presence of an Air–Water Interface Affects Formation and Elongation of α-Synuclein Fibrils

Cited by 251 publications

References 58 publications

Preparation and Characterization of Stable α-Synuclein Lipoprotein Particles

Preparation and Characterization of Stable α-Synuclein Lipoprotein Particles

α-Synuclein Insertion into Supported Lipid Bilayers As Seen by in Situ X-ray Reflectivity

Solution conditions determine the relative importance of nucleation and growth processes in α-synuclein aggregation

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