Issue 28, 2020, Issue in Progress
From the journal:

RSC Advances

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

Travis J. Nelson,‡a   Tiffany Truong,‡§a   BaoLong Truong,a   Camden V. Bilyeu,a   Jia Zhaoa  and  Cliff I. Stains ORCID logo *abcde  

* Corresponding authors

a Department of Chemistry, University of Nebraska – Lincoln, Lincoln, NE 68588, USA
E-mail: cstains@virginia.edu

b Nebraska Center for Integrated Biomolecular Communication, University of Nebraska – Lincoln, Lincoln, NE 68588, USA

c Cancer Genes and Molecular Regulation Program, Fred & Pamela Buffet Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA

d Department of Chemistry, University of Virginia, Charlottesville, VA, USA

e University of Virginia Cancer Center, University of Virginia, Charlottesville, VA, USA

Abstract

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

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

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Article information

DOI
https://doi.org/10.1039/D0RA02720K
Article type
Paper
Submitted
24 Mar 2020
Accepted
20 Apr 2020
First published
28 Apr 2020
This article is Open Access
Creative Commons BY-NC license

RSC Adv., 2020,10, 16675-16678

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

T. J. Nelson, T. Truong, B. Truong, C. V. Bilyeu, J. Zhao and C. I. Stains, RSC Adv., 2020, 10, 16675 DOI: 10.1039/D0RA02720K

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