Optimizing AB42-NR-Sup35M Protein Expression and Aggregation to Better Understand Alzheimer's Disease
Author(s)
Li, Julianne
Advisor(s)
Editor(s)
Collections
Supplementary to:
Permanent Link
Abstract
The general and overarching goal of this project is to better understand how amyloid formation contributes towards the progression and physiology of Alzheimer’s disease (AD). A prion is a type of misfolded protein and is typically composed of amyloids, or highly ordered across beta fibrils. Prions and amyloids are of particular interest to researchers because they are associated with over 50 human diseases and disorders, including neurodegenerative diseases like Creutzfeldt Jakob disease, Alzheimer’s, Parkinson’s, and Huntington’s disease. Of these neurodegenerative diseases and other forms of dementia, AD is the most common type in the world, with around 6.9 million individuals 65 and older in the United States alone living with the disease. Previous research has shown that the development of AD is closely related to the accumulation of amyloid beta, or Aβ, in the brain. However, the exact structure of these aggregations and how they contribute towards the condition is not completely understood by researchers. Thus, this project aims to better understand these aggregations. We optimized expression and purification of our Aβ42-NR-Sup35-M fusion construct in Escherichia coli cells before utilizing Saccharomyces cerevisiae yeast models to attempt in-vitro amyloid formation. We then performed Thioflavin-T (ThT) binding assays to assess amyloid formation. However, despite what we expected, our ThT assay results did not confirm any amyloid formation or aggregation of our Aβ42 fusion proteins. We will continue to manipulate our plasmid constructs and the environmental conditions in which we induced aggregation in attempts to successfully yield in vitro amyloid aggregation. Once optimized, our yeast model is a more affordable, equitable, and ethical alternative to using human or mammalian tissue to study amyloids and AD. By better understanding amyloid formation in AD, we can also better understand other prion disorders and bridge the gaps in research preventing researchers from developing an effective treatment or cure for AD.
Sponsor
Date
Extent
Resource Type
Text
Resource Subtype
Undergraduate Research Option Thesis