RT Journal Article
SR Electronic
T1 Label-free characterisation of amyloids and alpha-Synuclein polymorphs by exploiting their intrinsic fluorescence property
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.11.30.470691
DO 10.1101/2021.11.30.470691
A1 Chyi Wei Chung
A1 Amberley D. Stephens
A1 Edward Ward
A1 Yuqing Feng
A1 Molly Jo Davis
A1 Clemens F. Kaminski
A1 Gabriele S. Kaminski Schierle
YR 2021
UL http://biorxiv.org/content/early/2021/12/01/2021.11.30.470691.abstract
AB 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 represented by model-free phasor plots, as a label-free assay to characterise amyloid structure. Intrinsic amyloid fluorescence arises from 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 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 pre-formed fibrils of αS and βLG leads to increased fluorescence lifetimes, distinct to those of their fibrillar counterpart. 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 pathology), and for testing small molecule inhibitory compounds.Competing Interest StatementThe authors have declared no competing interest.