A luminescence-based assay for monitoring changes in alpha-synuclein aggregation in living cells. Issue 28 (28th April 2020)

• Record Type: Journal Article
• Authors: Nelson, Travis J.; Truong, Tiffany; Truong, BaoLong; Bilyeu, Camden V.; Zhao, Jia; Stains, Cliff I.
• Is Part Of: RSC advances. Volume 10:Issue 28(2020)
• Page Start: 16675
• Page End: 16678
• Publication Date: 2020-04-28
• Subjects: Chemistry -- Periodicals; 540.5
• DOI: 10.1039/d0ra02720k
• Languages: English
• ISSNs: 2046-2069

Abstract

Abstract : A re-engineered split-Nluc assay for detecting the influence of mutations and inhibitors on alpha-synuclein aggregation in 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 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.