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Ligand Profiling as a Diagnostic Tool to Differentiate Patient-Derived α-Synuclein Polymorphs

Published version
Peer-reviewed

Repository DOI


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Authors

paragon-plus: 6598372  ORCID logo  https://orcid.org/0000-0002-8693-3797
Melki, Ronald; paragon-plus: 6958201 
paragon-plus: 2703352  ORCID logo  https://orcid.org/0000-0002-5182-1859

Abstract

Amyloid fibrils are characteristic of many neurodegenerative diseases, including Alzheimer’s and Parkinson’s diseases. While different diseases may have fibrils formed of the same protein, the supramolecular morphology of these fibrils is disease-specific. Here, a method is reported to distinguish eight morphologically distinct amyloid fibrils based on differences in ligand binding properties. Eight fibrillar polymorphs of α-synuclein (αSyn) were investigated: five generated de novo using recombinant αSyn and three generated using protein misfolding cyclic amplification (PMCA) of recombinant αSyn seeded with brain homogenates from deceased patients diagnosed with Parkinson’s disease (PD), multiple system atrophy (MSA), and dementia with Lewy bodies (DLB). Fluorescence binding assays were carried out for each fibril using a toolkit of six different ligands. The fibril samples were separated into five categories based on a binary classification of whether they bound specific ligands or not. Quantitative binding measurements then allowed every fibrillar polymorph to be uniquely identified, and the PMCA fibrils derived from PD, MSA, and DLB patients could be unambiguously distinguished. This approach constitutes a novel and operationally simple method to differentiate amyloid fibril morphologies and to identify disease states using PMCA fibrils obtained by seeding with patient samples.

Description

Publication status: Published

Keywords

ligand binding site, Parkinson’s disease, polymorphs, protein misfolding cyclic amplification, fluorescence binding assay, α-synuclein

Journal Title

ACS Chemical Neuroscience

Conference Name

Journal ISSN

1948-7193
1948-7193

Volume Title

15

Publisher

American Chemical Society
Sponsorship
Equipment Call (EP/P030467/ , Engineering and Physical Sciences Research Council (Underpinning)
Cambridge Trust Prince of Wales Scholarship (NA)