1. Neuroscience
  2. Structural Biology and Molecular Biophysics
Download icon

An engineered monomer binding-protein for α-synuclein efficiently inhibits the proliferation of amyloid fibrils

  1. Emil Dandanell Agerschou
  2. Patrick Flagmeier
  3. Theodora Saridaki
  4. Céline Galvagnion
  5. Daniel Komnig
  6. Laetitia Heid
  7. Vibha Prasad
  8. Hamed Shaykhalishahi
  9. Dieter Willbold
  10. Christopher M Dobson
  11. Aaron Voigt
  12. Björn Falkenbürger  Is a corresponding author
  13. Wolfgang Hoyer  Is a corresponding author
  14. Alexander K Buell  Is a corresponding author
  1. Heinrich Heine University Düsseldorf, Germany
  2. University of Cambridge, United Kingdom
  3. RWTH Aachen University, Germany
  4. Technical University of Denmark, Denmark
Research Article
  • Cited 20
  • Views 3,762
  • Annotations
Cite this article as: eLife 2019;8:e46112 doi: 10.7554/eLife.46112

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

The following data sets were generated

Article and author information

Author details

  1. Emil Dandanell Agerschou

    Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Patrick Flagmeier

    Department of Chemistry, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1204-5340
  3. Theodora Saridaki

    Department of Neurology, RWTH Aachen University, Aachen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Céline Galvagnion

    Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Daniel Komnig

    Department of Neurology, RWTH Aachen University, Aachen, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6312-5236
  6. Laetitia Heid

    Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Vibha Prasad

    Department of Neurology, RWTH Aachen University, Aachen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Hamed Shaykhalishahi

    Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. Dieter Willbold

    Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0065-7366
  10. Christopher M Dobson

    Department of Chemistry, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  11. Aaron Voigt

    Department of Neurology, RWTH Aachen University, Aachen, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0428-7462
  12. Björn Falkenbürger

    Department of Neurology, RWTH Aachen University, Aachen, Germany
    For correspondence
    bfalkenburger@ukaachen.de
    Competing interests
    The authors declare that no competing interests exist.
  13. Wolfgang Hoyer

    Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
    For correspondence
    wolfgang.hoyer@hhu.de
    Competing interests
    The authors declare that no competing interests exist.
  14. Alexander K Buell

    Biotechnology and Biomedicine, Technical University of Denmark, Kgs Lyngby, Denmark
    For correspondence
    alebu@dtu.dk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1161-3622

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

  1. Andrew B West, Duke University, United States

Publication history

  1. Received: February 25, 2019
  2. Accepted: August 4, 2019
  3. Accepted Manuscript published: August 7, 2019 (version 1)
  4. Accepted Manuscript updated: August 21, 2019 (version 2)
  5. Version of Record published: September 3, 2019 (version 3)
  6. 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

  • 3,762
    Page views
  • 610
    Downloads
  • 20
    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)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

Further reading

    1. Neuroscience
    Kara A Fulton, Kevin L Briggman
    Tools and Resources

    A dense reconstruction of neuronal synaptic connectivity typically requires high-resolution 3D electron microscopy (EM) data, but EM data alone lacks functional information about neurons and synapses. One approach to augment structural EM datasets is with the fluorescent immunohistochemical (IHC) localization of functionally relevant proteins. We describe a protocol that obviates the requirement of tissue permeabilization in thick tissue sections, a major impediment for correlative pre-embedding IHC and EM. We demonstrate the permeabilization-free labeling of neuronal cell types, intracellular enzymes, and synaptic proteins in tissue sections hundreds of microns thick in multiple brain regions from mice while simultaneously retaining the ultrastructural integrity of the tissue. Finally, we explore the utility of this protocol by performing proof-of-principle correlative experiments combining two-photon imaging of protein distributions and 3D EM.

    1. Neuroscience
    Alexa Pichet Binette et al.
    Research Article

    Beta-amyloid (Aβ) and tau proteins, the pathological hallmarks of Alzheimer's disease (AD), are believed to spread through connected regions of the brain. Combining diffusion imaging and positron emission tomography, we investigated associations between white matter microstructure specifically in bundles connecting regions where Aβ or tau accumulates and pathology. We focussed on free-water corrected diffusion measures in the anterior cingulum, posterior cingulum, and uncinate fasciculus in cognitively normal older adults at risk of sporadic AD and presymptomatic mutation carriers of autosomal dominant AD. In Aβ-positive or tau-positive groups, lower tissue fractional anisotropy and higher mean diffusivity related to greater Aβ and tau burden in both cohorts. Associations were found in the posterior cingulum and uncinate fasciculus in preclinical sporadic AD, and in the anterior and posterior cingulum in presymptomatic mutation carriers. These results suggest that microstructural alterations accompany pathological accumulation as early as the preclinical stage of both sporadic and autosomal dominant AD.