Alpha-synuclein (ASYN) is considered an intrinsically disordered protein and constitutes the major protein component of amyloid aggregates associated with Parkinson's disease (PD). Given the clear role of ASYN aggregation in PD, there is a critical need for assays capable of reporting on changes in the solubility of ASYN in living systems. We have recently discovered self-assembling fragments of NanoLuc luciferase (Nluc) that can be utilized as probes of protein solubility within living cells. In this system, self-assembly of Nluc fragments is proportional to the solubility of a protein fused to the N-terminus of the N-terminal Nluc fragment, providing a turn-on signal for increased protein solubility. Utilizing this approach allows for the investigation of the effect of small molecules, as well as mutations on protein solubility within living systems. Herein, we describe the development of an assay system capable of reporting on changes in ASYN solubility within living cells. We demonstrate the ability to detect changes in ASYN aggregation potential due to mutations as well as small molecules. In the long term, this platform could be used to provide insight into the structural determinants of ASYN aggregation as well as identify inhibitors of ASYN aggregation in living cells.

Self-Assembling nanoluciferase fragments as probes for alpha-synuclein solubility

253rd National Meeting (2017)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.