Label-Free Characterization of Amyloids and Alpha-Synuclein Polymorphs by Exploiting Their Intrinsic Fluorescence Property.
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Authors
Ward, Edward
Feng, Yuqing
Davis, Molly Jo
Publication Date
2022-04-05Journal Title
Anal Chem
ISSN
0003-2700
Publisher
American Chemical Society (ACS)
Volume
94
Issue
13
Pages
5367-5374
Language
eng
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Chung, C. W., Stephens, A. D., Ward, E., Feng, Y., Davis, M. J., Kaminski, C. F., & Kaminski Schierle, G. S. (2022). Label-Free Characterization of Amyloids and Alpha-Synuclein Polymorphs by Exploiting Their Intrinsic Fluorescence Property.. Anal Chem, 94 (13), 5367-5374. https://doi.org/10.1021/acs.analchem.1c05651
Description
Funder: Infinitus China Ltd.
Abstract
Conventional in vitro aggregation assays often involve tagging with extrinsic fluorophores, which can interfere with aggregation. We propose the use of intrinsic amyloid fluorescence lifetime probed using two-photon excitation and represented by model-free phasor plots as a label-free assay to characterize the amyloid structure. Intrinsic amyloid fluorescence arises from the structured packing of β-sheets in amyloids and is independent of aromatic-based fluorescence. We show that different amyloids [i.e., α-Synuclein (αS), β-Lactoglobulin (βLG), and TasA] and different polymorphic populations of αS (induced by aggregation in salt-free and salt buffers mimicking the intra-/extracellular environments) can be differentiated by their unique fluorescence lifetimes. Moreover, we observe that disaggregation of the preformed fibrils of αS and βLG leads to increased fluorescence lifetimes, distinct from those of their fibrillar counterparts. Our assay presents a medium-throughput method for rapid classification of amyloids and their polymorphs (the latter of which recent studies have shown lead to different disease pathologies) and for testing small-molecule inhibitory compounds.
Keywords
Amyloid, Fluorescence, alpha-Synuclein, Amyloidogenic Proteins, Protein Conformation, beta-Strand
Relationships
Is supplemented by: https://doi.org/10.17863/CAM.82267
Sponsorship
Medical Research Council (MR/K02292X/1)
Wellcome Trust (065807/Z/01/Z)
Wellcome Trust (203249/Z/16/Z)
Engineering and Physical Sciences Research Council (EP/S022953/1)
Identifiers
35333515, PMC8988127
External DOI: https://doi.org/10.1021/acs.analchem.1c05651
This record's URL: https://www.repository.cam.ac.uk/handle/1810/336436
Rights
Attribution-NonCommercial-NoDerivatives 4.0 International
Licence URL: https://creativecommons.org/licenses/by-nc-nd/4.0/
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