Alpha-synuclein is a 14kD protein that is strongly associated with Parkinson’s disease. This protein is a major component of Lewy Bodies, which are a distinctive feature of Parkinson’s. Alpha-synuclein exists as an intrinsically disordered protein and when it forms fibrils it has shown to be toxic to cells. Examining alpha-synuclein <i>in vitro</i> has given further insight into how this protein forms aggregates, but the mechanism of cell death is still unknown. Different constructs of alpha-synuclein can be labeled at the N and C termini with Tetramethylrhodamine, a relatively small fluorophore, which could decrease the chance of hindering the aggregation properties of alpha-synuclein. Characteristics of purified alpha-synuclein constructs can be distinguished using Circular Dichroism, Fourier Transform Infrared Spectroscopy, Transmission Electron Microscopy, and Aggregation studies of turbidity and ThT response over time. Different cell lines have varying levels of endogenous alpha-synuclein. By treating different cell lines with the different polymorphs of alpha-synuclein and examining the differing effects the fibrils have on the cell lines, a further understanding can be attained about endogenous levels of this protein contributing to the survival of the cells. Observable differences in fluorescence may also illuminate how the protein changes structurally when the fibrils internalize in the cell. By observing where these fibrils localize in cells, visualizing the morphological differences, and coupling to chemical viability assays, we hope to gain a further understanding of the cell-killing mechanism.

Effects of Alpha-synuclein uptake on cellular viability, morphology, and localization

254th National Meeting (2017)

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