Year: 2016
Type: dissertation
Abstract: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the loss of motor neurons resulting in progressive paralysis that eventually leads to the death of an afflicted individual. In a majority of ALS cases, the cause of onset is unknown, but a small subset of ALS cases are linked to mutations in the protein Cu, Zn superoxide dismutase (SOD1). Despite extensive research in this area, few treatments for ALS have been developed. Since the description of the disease by Jean-Martin Charcot in the mid-nineteenth century, only one drug, Riluzole has been approved in the management of this disease. We propose investigating therapeutic treatments of ALS by looking for small molecules that have the potential to stabilize the native structure of mutant SOD1 and prevent its misfolding and aggregation. We started our investigation by first devising a method to optimize the purification of wildtype and mutant SOD1 in our lab. We constructed a fusion protein of SOD1 linked to a yellow fluorescent protein, a hexahistidine tag, and a tobacco etch virus (TEV) cleavage site and showed that both wildtype and mutant SOD1 purified from this construct was active and metallated. We used our purified protein to explore the results of an in silico binding screen searching for SOD1 native-state binders. We discovered the flavonoid, epigallocatecin-3-gallate (EGCG) was able to inhibit the misfolding and aggregation of the ALS-causing mutation A4V-SOD1 exposed to oxidative stress. Additionally, we have evidence that the inhibitory effect of EGCG occurs through the stabilization of the dimer interface of mutant SOD1, rather than the anti-oxidant properties of this molecule. This is the first report of EGCG having anti-aggregation properties in an ALS model and these results implicate the potential therapeutic value of EGCG within the field of SOD1-ALS.
Source: TSpace (University of Toronto)
Author: Priya Sharda
Language: German
Subfield: Clinical Neurology
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W2561503337