Date

5-7-2024

Document Type

Thesis

Abstract

Mitoxantrone is a chemotherapy drug that treats a variety of cancers by inhibiting cancer cell replication through intercalation with DNA. Intercalation is the process by which certain small molecules slide flat planar regions in their structure between the base pairs of DNA. This increases the stacking force in the DNA double helix, preventing DNA replication by increasing the force required to separate the two strands that compose the helix. In this study, dual-beam optical tweezers were used to determine the binding kinetics and dissociation constant of Mitoxantrone to single molecules of DNA at 20 pN of force. This study utilized a method known as constant force experiments. During these experiments, a single DNA molecule was trapped and stretched to a specified force. The DNA molecule was held at this force, with the assistance of a force feedback program, while Mitoxantrone was introduced to the system and allowed to reach equilibrium binding with DNA. We found an off rate of 0.132 ± 0.018 s-1, an association rate of 10.5 ± 0.7 μM-1s-1, and the dissociation constant of 11.3 ± 0.8 nM for MTX. Our results show that MTX binds to DNA in the nanomolar range, in contrast to the micromolar range reported in the literature. This may prompt further studies on MTX dosage in the future.

Department

Physics, Photonics and Optical Engineering

Thesis Comittee

Dr. Thayaparan Paramanathan, Thesis Advisor
Dr. Edward Deveney, Committee Member
Dr. Thomas Kling, Committee Member

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