A luminescence-based assay for monitoring changes in alpha-synuclein aggregation in living cells

Travis J. Nelson; Tiffany Truong; Bao Long Truong; Camden V. Bilyeu; Jia Zhao; Cliff I. Stains

doi:10.1039/d0ra02720k

A luminescence-based assay for monitoring changes in alpha-synuclein aggregation in living cells

Travis J. Nelson, Tiffany Truong, Bao Long Truong, Camden V. Bilyeu, Jia Zhao, Cliff I. Stains

Research output: Contribution to journal › Article › peer-review

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Abstract

Parkinson's disease is characterized by the accumulation of protein aggregates in the brain, termed Lewy bodies. Lewy bodies are predominantly composed of a-synuclein and mutations that increase the aggregation potential of a-synuclein have been associated with early on-set disease. Assays capable of reporting on the solubility of a-synuclein in living cells could provide a means to interrogate the influence of mutations on aggregation as well as identify small molecules capable of modulating the aggregation of a-synuclein. Herein, we repurpose our previously reported self-assembling NanoLuc luciferase fragments to engineer a platform for detecting a-synuclein solubility in living cells. This new assay is capable of reporting on changes in a-synuclein solubility caused by disease-relevant mutations as well as inhibitors of aggregation. In the long term, this new assay platform provides a means to investigate the influence of mutations on a-synuclein solubility as well as identify potential tool compounds capable of modulating a-synuclein aggregation.

Original language	English (US)
Pages (from-to)	16675-16678
Number of pages	4
Journal	RSC Advances
Volume	10
Issue number	28
DOIs	https://doi.org/10.1039/d0ra02720k
State	Published - Apr 28 2020

ASJC Scopus subject areas

Chemistry(all)
Chemical Engineering(all)

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title = "A luminescence-based assay for monitoring changes in alpha-synuclein aggregation in living cells",

abstract = "Parkinson's disease is characterized by the accumulation of protein aggregates in the brain, termed Lewy bodies. Lewy bodies are predominantly composed of a-synuclein and mutations that increase the aggregation potential of a-synuclein have been associated with early on-set disease. Assays capable of reporting on the solubility of a-synuclein in living cells could provide a means to interrogate the influence of mutations on aggregation as well as identify small molecules capable of modulating the aggregation of a-synuclein. Herein, we repurpose our previously reported self-assembling NanoLuc luciferase fragments to engineer a platform for detecting a-synuclein solubility in living cells. This new assay is capable of reporting on changes in a-synuclein solubility caused by disease-relevant mutations as well as inhibitors of aggregation. In the long term, this new assay platform provides a means to investigate the influence of mutations on a-synuclein solubility as well as identify potential tool compounds capable of modulating a-synuclein aggregation.",

author = "Nelson, {Travis J.} and Tiffany Truong and Truong, {Bao Long} and Bilyeu, {Camden V.} and Jia Zhao and Stains, {Cliff I.}",

note = "Funding Information: This work was funded by the NIH (R35GM119751) and the University of Virginia. The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. Publisher Copyright: {\textcopyright} The Royal Society of Chemistry 2020.",

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AU - Truong, Tiffany

AU - Truong, Bao Long

AU - Bilyeu, Camden V.

AU - Zhao, Jia

AU - Stains, Cliff I.

N1 - Funding Information: This work was funded by the NIH (R35GM119751) and the University of Virginia. The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. Publisher Copyright: © The Royal Society of Chemistry 2020.

PY - 2020/4/28

Y1 - 2020/4/28

N2 - Parkinson's disease is characterized by the accumulation of protein aggregates in the brain, termed Lewy bodies. Lewy bodies are predominantly composed of a-synuclein and mutations that increase the aggregation potential of a-synuclein have been associated with early on-set disease. Assays capable of reporting on the solubility of a-synuclein in living cells could provide a means to interrogate the influence of mutations on aggregation as well as identify small molecules capable of modulating the aggregation of a-synuclein. Herein, we repurpose our previously reported self-assembling NanoLuc luciferase fragments to engineer a platform for detecting a-synuclein solubility in living cells. This new assay is capable of reporting on changes in a-synuclein solubility caused by disease-relevant mutations as well as inhibitors of aggregation. In the long term, this new assay platform provides a means to investigate the influence of mutations on a-synuclein solubility as well as identify potential tool compounds capable of modulating a-synuclein aggregation.

AB - Parkinson's disease is characterized by the accumulation of protein aggregates in the brain, termed Lewy bodies. Lewy bodies are predominantly composed of a-synuclein and mutations that increase the aggregation potential of a-synuclein have been associated with early on-set disease. Assays capable of reporting on the solubility of a-synuclein in living cells could provide a means to interrogate the influence of mutations on aggregation as well as identify small molecules capable of modulating the aggregation of a-synuclein. Herein, we repurpose our previously reported self-assembling NanoLuc luciferase fragments to engineer a platform for detecting a-synuclein solubility in living cells. This new assay is capable of reporting on changes in a-synuclein solubility caused by disease-relevant mutations as well as inhibitors of aggregation. In the long term, this new assay platform provides a means to investigate the influence of mutations on a-synuclein solubility as well as identify potential tool compounds capable of modulating a-synuclein aggregation.

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A luminescence-based assay for monitoring changes in alpha-synuclein aggregation in living cells

Abstract

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