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Open AccessArticle

Kinetic Transition in Amyloid Assembly as a Screening Assay for Oligomer-Selective Dyes

Department of Physics, University of South Florida, Tampa, FL 33620, USA
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Author to whom correspondence should be addressed.
Biomolecules 2019, 9(10), 539; https://doi.org/10.3390/biom9100539
Received: 6 September 2019 / Revised: 24 September 2019 / Accepted: 25 September 2019 / Published: 27 September 2019
(This article belongs to the Special Issue 2019 Feature Papers by Biomolecules’ Editorial Board Members)
Assembly of amyloid fibrils and small globular oligomers is associated with a significant number of human disorders that include Alzheimer’s disease, senile systemic amyloidosis, and type II diabetes. Recent findings implicate small amyloid oligomers as the dominant aggregate species mediating the toxic effects in these disorders. However, validation of this hypothesis has been hampered by the dearth of experimental techniques to detect, quantify, and discriminate oligomeric intermediates from late-stage fibrils, in vitro and in vivo. We have shown that the onset of significant oligomer formation is associated with a transition in thioflavin T kinetics from sigmoidal to biphasic kinetics. Here we showed that this transition can be exploited for screening fluorophores for preferential responses to oligomer over fibril formation. This assay identified crystal violet as a strongly selective oligomer-indicator dye for lysozyme. Simultaneous recordings of amyloid kinetics with thioflavin T and crystal violet enabled us to separate the combined signals into their underlying oligomeric and fibrillar components. We provided further evidence that this screening assay could be extended to amyloid-β peptides under physiological conditions. Identification of oligomer-selective dyes not only holds the promise of biomedical applications but provides new approaches for unraveling the mechanisms underlying oligomer versus fibril formation in amyloid assembly. View Full-Text
Keywords: oligomer formation; amyloidosis; assembly pathway; dye fluorescence; thioflavin T oligomer formation; amyloidosis; assembly pathway; dye fluorescence; thioflavin T
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MDPI and ACS Style

Barton, J.; Arias, D.S.; Niyangoda, C.; Borjas, G.; Le, N.; Mohamed, S.; Muschol, M. Kinetic Transition in Amyloid Assembly as a Screening Assay for Oligomer-Selective Dyes. Biomolecules 2019, 9, 539.

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