Study of α-Synuclein Pathologic Effect at Cellular and Mitochondrial Level (2010)

Undergraduates: Yoo Jeong Yang, none Imola G. Zigoneanu none

Faculty Advisor: Gary Pielak
Department: Chemistry

---

α-Synuclein is a natively disordered neuron specific protein that has implications in Parkinson’s disease. Recent studies suggest α-synuclein localization in mitochondria and toxicity of α-synuclein protofibrils as a primary cause of the neurodegenerative diseases. The overarching goal of this research is to study these two aspects more in depth to give a better insight into the mechanism of progression of Parkinson’s disease at cellular level. α-synuclein was translocated into the isolated mitochondria of mammalian cells to understand localization of α-synuclein inside the cells. This was achieved by incubating freshly isolated mitochondria from HeLa cells with fluorescently labeled V3C α-synuclein followed by determination of protein localization in different mitochondria compartments. α-Synuclein was found in the intermembrane space and inner membrane of mitochondria, indicating possible localization of α-synuclein in cells in Parkinson’s disease. Also, protofibril species of α-synuclein was synthesized in vitro to test their effect on neuronal viability. Wild type α-synuclein, overexpressed in E.coli and purified using FPLC, was used for synthesis of spherical protofibrils. This was confirmed by transmission electron microscopy. The protofibrils will be microinjected into primary culture neurons dissected from superior cervical ganglia of mice. It is expected that α-synuclein protofibrils will affect neuronal viability at higher rate compared to monomeric proteins.