Selectivity of Lewy body protein interactions along the aggregation pathway of α-synuclein

André D G Leitão; Paulina Rudolffi-Soto; Alexandre Chappard; Akshay Bhumkar; Derrick Lau; Dominic J B Hunter; Yann Gambin; Emma Sierecki

doi:10.1038/s42003-021-02624-x

Selectivity of Lewy body protein interactions along the aggregation pathway of α-synuclein

Commun Biol. 2021 Sep 23;4(1):1124. doi: 10.1038/s42003-021-02624-x.

Authors

André D G Leitão^{1

2}, Paulina Rudolffi-Soto¹, Alexandre Chappard^{1

3}, Akshay Bhumkar^{1

4}, Derrick Lau¹, Dominic J B Hunter^{1

5}, Yann Gambin⁶, Emma Sierecki⁷

Affiliations

¹ EMBL Australia Node in Single Molecule Science and School of Medical Sciences, The University of New South Wales, Sydney, NSW, Australia.
² Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA, USA.
³ School of Chemistry, The University of Edinburgh, Edinburgh, UK.
⁴ Woolcock Institute of Medical Research, University of Sydney, Sydney, NSW, Australia.
⁵ Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, Australia.
⁶ EMBL Australia Node in Single Molecule Science and School of Medical Sciences, The University of New South Wales, Sydney, NSW, Australia. y.gambin@unsw.edu.au.
⁷ EMBL Australia Node in Single Molecule Science and School of Medical Sciences, The University of New South Wales, Sydney, NSW, Australia. e.sierecki@unsw.edu.au.

Abstract

The aggregation of alpha-synuclein (α-SYN) follows a cascade of oligomeric, prefibrillar and fibrillar forms, culminating in the formation of Lewy Bodies (LB), the pathological hallmarks of Parkinson's Disease. Although LB contain over 70 proteins, the potential for interactions along the aggregation pathway of α-SYN is unknown. Here we propose a map of interactions of 65 proteins against different species of α-SYN. We measured binding to monomeric α-SYN using AlphaScreen, a sensitive nano-bead luminescence assay for detection of protein interactions. To access oligomeric species, we used the pathological mutants of α-SYN (A30P, G51D and A53T) which form oligomers with distinct properties. Finally, we generated amyloid fibrils from recombinant α-SYN. Binding to oligomers and fibrils was measured by two-color coincidence detection (TCCD) on a single molecule spectroscopy setup. Overall, we demonstrate that LB components are recruited to specific steps in the aggregation of α-SYN, uncovering future targets to modulate aggregation in synucleinopathies.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amyloid / metabolism
Humans
Lewy Bodies / metabolism*
Parkinson Disease / pathology*
alpha-Synuclein / metabolism*

Substances

Amyloid
SNCA protein, human
alpha-Synuclein