Date of Graduation

5-2022

Document Type

Thesis

Degree Name

Master of Science in Cell & Molecular Biology (MS)

Degree Level

Graduate

Department

Biological Sciences

Advisor/Mentor

Chenguang Fan

Committee Member

Josh Sakon

Second Committee Member

Yuchun Du

Keywords

Aconitase, Enzyme dysfunction, Genetic Code Expansion, Kreb's cycle, Mitochondria, TCA Cycle

Abstract

The tricarboxylic acid (TCA) cycle is a very important, centrally located, energy-producing pathway that connects numerous other metabolic and regulatory pathways. Enzymes of this cycle have been more recently implicated in various cancers and neurometabolic disorders, however, the exact mechanism by which this happens becomes quite complex when considering the potential modification of these enzymes and the presence of multiple forms of the enzymes and therefore there is much to be studied in this area.

Aconitase has become a recent enzyme of interest as its substrate, citrate, has been found to play a major role in many vital processes within an organism, including the survival and expansion of cancer. Additionally, the modification of aconitase by acetylation has been recently found to drive its activation to support energy, growth, and metastasis in prostate cancer tissue. In this master’s thesis, I apply the genetic code expansion technique to E. coli aconitase isozymes AcnA and AcnB to examine the changes in function caused by the modification of the structure in order to achieve a greater understanding of the impact of lysine acetylation in these isozymes.

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