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

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