 | Marisa Rubio Marshall University (Chemistry '08)Mentor(s)David Talaga, Ph.D. Cheng-Yen Huang, Ph.D. Jason Giurleo Department of Chemistry and Chemical Biology Rutgers University |
| ||
| Characterization of amyloid fibril aggregation by single molecule fluorescence | ||
| Several neurodegenerative pathologies such as Alzheimer’s and Parkinson’s diseases are characterized by amyloid fibril protein build-up in the brain. However, the formation and aggregation of these thread-like protein fibrils is not well understood. The negative effects of these diseases have provided impetus for elucidating the mechanism of amyloid fibril formation. This in vitro study uses single molecule fluorescence confocal microscopy to characterize the protein precursors along the amyloid pathway, providing insight into the mechanism by which amyloid fibrils form. This experiment allows for a level of species classification difficult to accomplish in bulk (ensemble averaged) measurements. 200mM a-synuclein, the protein implicated in Parkinson’s disease, was labeled with the environmentally sensitive fluorophore Nile Red and incubated at 15°C for 8 days. Varying lifetimes were seen for monomers, aggregates, and fibrils of 2-4, 4-6, and 0.4-2 ns, respectively, due to the unique environment each conformation creates. These lifetime-species correlations through single molecule fluorescence provide more certain identification of species in bulk measurements. |