An engineered monomer binding-protein for α-synuclein efficiently inhibits the proliferation of amyloid fibrils
- Heinrich Heine University Düsseldorf, Germany
- University of Cambridge, United Kingdom
- RWTH Aachen University, Germany
- Technical University of Denmark, Denmark
Abstract
Removing or preventing the formation of α-synuclein aggregates is a plausible strategy against Parkinson's disease. To this end we have engineered the β-wrapin AS69 to bind monomeric α-synuclein with high affinity. In cultured cells, AS69 reduced the self-interaction of α-synuclein and the formation of visible α-synuclein aggregates. In flies, AS69 reduced α-synuclein aggregates and the locomotor deficit resulting from α-synuclein expression in neuronal cells. In biophysical experiments in vitro, AS69 highly sub-stoichiometrically inhibited both primary and autocatalytic secondary nucleation processes, even in the presence of a large excess of monomer. We present evidence that the AS69-α-synuclein complex, rather than the free AS69, is the inhibitory species responsible for sub-stoichiometric inhibition of secondary nucleation. These results represent a new paradigm that high affinity monomer binders can lead to strongly sub-stoichiometric inhibition of nucleation processes.
Data availability
- Numerical data represented in the graphs for cell culture and fly experiments will be made publicly available on osf.io as we did for previous publications.- The numerical data for the biophysical experiments will be made publicly available within the same repository on osf.io.- The raw images of the gels used in the publication will be made publicly available.All data have been deposited on osf.io ( https://osf.io/6n2gs/?view_only=7eb7024d8ecb460a817cd0ed35978339 ) and will be made available in the event of publication
Article and author information
Author details
Funding
Leverhulme Trust
- Alexander K Buell
Boehringer Ingelheim Fonds
- Patrick Flagmeier
Studienstiftung des Deutschen Volkes
- Patrick Flagmeier
Alexander von Humboldt-Stiftung
- Céline Galvagnion
H2020 European Research Council (726368)
- Céline Galvagnion
Parkinson's and Movement Disorder Foundation
- Alexander K Buell
European Commission (706551)
- Céline Galvagnion
Novo Nordisk Foundation
- Alexander K Buell
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Andrew B West, Duke University, United States
Version history
- Received: February 25, 2019
- Accepted: August 4, 2019
- Accepted Manuscript published: August 7, 2019 (version 1)
- Accepted Manuscript updated: August 21, 2019 (version 2)
- Version of Record published: September 3, 2019 (version 3)
- Version of Record updated: December 11, 2019 (version 4)
Copyright
© 2019, Agerschou et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 5,142
- Page views
-
- 809
- Downloads
-
- 43
- Citations
Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, PubMed Central.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
An engineered monomer binding-protein for α-synuclein efficiently inhibits the proliferation of amyloid fibrils
eLife 8:e46112.
https://doi.org/10.7554/eLife.46112
Further reading
-
- Chromosomes and Gene Expression
- Neuroscience
Mathys et al. conducted the first single-nucleus RNA-seq (snRNA-seq) study of Alzheimer’s disease (AD) (Mathys et al., 2019). With bulk RNA-seq, changes in gene expression across cell types can be lost, potentially masking the differentially expressed genes (DEGs) across different cell types. Through the use of single-cell techniques, the authors benefitted from increased resolution with the potential to uncover cell type-specific DEGs in AD for the first time. However, there were limitations in both their data processing and quality control and their differential expression analysis. Here, we correct these issues and use best-practice approaches to snRNA-seq differential expression, resulting in 549 times fewer DEGs at a false discovery rate of 0.05. Thus, this study highlights the impact of quality control and differential analysis methods on the discovery of disease-associated genes and aims to refocus the AD research field away from spuriously identified genes.
-
- Neuroscience
The strength of a fear memory significantly influences whether it drives adaptive or maladaptive behavior in the future. Yet, how mild and strong fear memories differ in underlying biology is not well understood. We hypothesized that this distinction may not be exclusively the result of changes within specific brain regions, but rather the outcome of collective changes in connectivity across multiple regions within the neural network. To test this, rats were fear conditioned in protocols of varying intensities to generate mild or strong memories. Neuronal activation driven by recall was measured using c-fos immunohistochemistry in 12 brain regions implicated in fear learning and memory. The interregional coordinated brain activity was computed and graph-based functional networks were generated to compare how mild and strong fear memories differ at the systems level. Our results show that mild fear recall is supported by a well-connected brain network with small-world properties in which the amygdala is well-positioned to be modulated by other regions. In contrast, this connectivity is disrupted in strong fear memories and the amygdala is isolated from other regions. These findings indicate that the neural systems underlying mild and strong fear memories differ, with implications for understanding and treating disorders of fear dysregulation.
