Date of Submission

Spring 2024

Academic Program

Biology; Chemistry and Biochemistry

Project Advisor 1

Swapan Jain

Project Advisor 2

Michael Tibbetts

Abstract/Artist's Statement

Cancer is the second leading cause of death in the United States, with an estimated 609,820 cancer-related deaths in 2023. In recent decades, researchers have tapped into the potential of metal-based compounds as anticancer chemotherapeutic agents. Researchers have discovered that adding metal centers to common drugs could increase efficacy by improving both activity and properties of inhibition. They have also investigated the possibility of producing drugs with synergistic effects that display potency similar to successful drugs such as Carboplatin and KP-1019, while simultaneously reducing known cytotoxicities. Dihydrofolate reductase (DHFR) is an enzyme involved in folate metabolism, which is an essential component of DNA synthesis. If metal-based compounds inhibit folate production through their effects on DHFR, this might limit DNA synthesis. In turn, cancer cells might not be able to proliferate, mitigating the growth of malignancies.

In this project, I aimed to understand the effects Ruthenium-based drugs have on the function of the DHFR enzyme compared to known inhibitors. The first step was to transform BL21-competent Escherichia coli cells with a plasmid to overexpress the DHFR enzyme, which could then be purified. To identify, quantify, and examine the purified protein, I performed Bradford assays, enzymatic activity assays, Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis (SDS-PAGE), and time and concentration affected induction assays. Although I intended to investigate the effects of Ruthenium-based drugs on the function of the DHFR enzyme, numerous obstacles prevented me from successfully answering the questions. Future directions include conducting further research to successfully obtain a pure protein and utilize techniques such as electrophoretic mobility shift assays (EMSA) and isothermal titration calorimetry to understand how Ruthenium-based drugs impact enzyme activity in comparison to known inhibitors and examine the binding affinity of the Ruthenium mononuclear complexes to the DHFR enzyme.

Open Access Agreement

On-Campus only

Creative Commons License

Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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