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Innovate Summer Experience: Examining alpha-Synuclein Nitration in Parkinson’s Disease

Yuliya Zayats 
Department of Biology 
Lake Forest College 
Lake Forest, IL 60045 

This summer, I conducted research in Dr. Shubhik DebBurman’s laboratory which explored the molecular biology of Parkinson’s disease, the second most common neurodegenerative disorder. Parkinson’s dis- ease pathology is characterized by the buildup of a misfolded protein called α-synuclein, which in turn forms insoluble aggregates called Lewy Bodies. As a result, Lewy Bodies cause the death of brain cells. Research on Par- kinson’s disease is crucial in finding successful treatments, for the disease impacts a large percentage of the elderly population. However, to properly design a treatment for Parkinson’s, one must first understand the properties of α-synuclein itself and how it can be altered to decrease or even prevent aggregation of the protein. I had the opportunity to conduct experiments that could potentially contribute to groundbreaking Parkinson’s treatments very early in my college career. The Richter research gave me an insight of what it is like being a scientist as well as how my studies in biology will help me develop a career that will have a significant impact on treatment of diseases.

My project focused on α-synuclein’s ability to be modified with the attachment of chemical groups. In particular, a modification known as nitration, which is an attachment of a nitro group onto a protein. The nitro group can be attached to four different sites on α-synuclein and Parkin- son’s patients exhibit the nitration of all four possible sites. In fact, a past senior thesis student in Dr. DebBurman’s lab, Keith Solvang, conducted re- search on α-synuclein that was modified only on one site. By the end of his research, Keith came to the conclusion that when α-synuclein is nitrated on only one site it can regulate the proteins’ properties, but only to modest degrees. Keith’s senior thesis led to my summer research, which explored α-synuclein nitration not only on one site, but on different combinations of sites. However, what was unknown was how individual nitrated sites contribute to α-synuclein properties and whether there was a specific com- bination that has the most impact on α-synuclein properties. I conjectured that the combination of nitration sites has the most impact on α-synuclein characteristics, as opposed to nitration on one site. To test my hypothesis, my goal was to create and characterize α-synuclein combination mutants, which are proteins that are mutated on numerous sites instead of one.

The strategy devised for creating the combination mutants was coherent, but executing the procedure proved to pose many challenges. This not only taught me many important lab techniques, but forced me to think critically about how I approach complications in executing exper- iments. My peers, as well as Dr. Shubhik DebBurman, helped me learn every step in the process as well as expand my knowledge of the lab- oratory. In completing my project, I worked alongside Charles Alvarado and Morgan Marshall with whom I carried out every experiment and who made the goal of creating combination mutants attainable. In our group we were able to work in collaboration, as well as with a lot of enthusiasm, which earned us our name “Nitration Squad”. Together we carried out nu- merous experiments such as green fluorescence protein (GFP) micros- copy, which gave us an insight as to where α-synuclein accumulated in the cells. We also performed many mutagenesis reactions that created the desired mutations of the proteins that could yield nitration at specific sites, as well as bacterial and yeast transformations. This made the place- ment of mutations and their expression possible in both bacteria and yeast. Although not all of our mutagenesis reactions and transformations were successful, we learned to pursue our research with determination when we faced issues and used logical reasoning to help us eliminate those issues.

My Richter scholar research has proven to be a unique experi- ence that expanded my knowledge and taught me many valuable skills. The variety of challenges that came with the project were not mishaps, but rather opportunities to become a more conscientious and knowledgeable researcher. Not only did the research in Dr. Deburman’s laboratory enable me to grow intellectually, but it contributed to the overall understanding of the characteristics of α-synuclein protein, which is the hallmark of Parkinson’s disease. The work done by Dr. DebBurman and Lake Forest undergraduate students is of great significance and I was very fortunate to be a part of it.


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