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Graduate Defense: Michael Blair

June 27, 2023 @ 2:00 pm - 3:00 pm

Thesis Defense

Thesis Information

Title: A Comparison of Methods Used to Isolate DNA Protein Crosslinks Formed by Aziridinomitosenes

Program: Master of Science in Chemistry

Advisor: Dr. Don Warner, Chemistry and Biochemistry

Committee Members: Dr. Michael Callahan, Chemistry and Biochemistry; and Dr. Ken Cornell, Chemistry and Biochemistry


Aziridinomitosenes are synthetic chemotherapeutic agents that were discovered from Mitomycin C, as well as other mitomycins. They are known for their ability to form DNA interstrand cross-links, and more recently have been observed to form DNA-protein cross-links. In this work, three synthetic aziridinomitsenes were prepared using a previously developed method to better understand the role of the C6 and C7 positions in the formation of DNA-protein cross-links. In addition to this, two separate methods for isolating and quantifying DNA-protein cross-links were utilized and compared to assess the advancements in the technique. H AZM, C6 AZM, Dimethyl AZM, and Mitomycin C were compared to formaldehyde, a known DNA-protein cross-linking agent in these techniques. Results from the ARK assay displayed a 4x increase in average response in nearly every treatment group when compared to the previously developed K-SDS method. Additionally, the ARK assay provided tighter standard deviations indicating more reliable data. Furthermore, analysis of ARK data displayed that the C6 AZM was capable of forming a greater percentage of DNA-protein cross-links compared to the H AZM and Dimethyl AZM. These results indicated that the C6 position of aziridinomitosenes may play a much more important role in DNA-protein cross-link formation.