Date of Graduation

5-2021

Document Type

Thesis

Degree Name

Master of Science in Cell & Molecular Biology (MS)

Degree Level

Graduate

Department

Biological Sciences

Advisor

Paul D. Adams

Committee Member

Matt McIntosh

Second Committee Member

Suresh Kumar Thallapuranam

Third Committee Member

Ines Pinto

Keywords

anticancer drug therapy, biophysical characterization, Gibbs free energy, fluorescence denaturation, molecular interactions

Abstract

Eukaryotic cells contain an extensive amount of GTP/GDP binding proteins. Proteins known as Ras GTPase primary function as a binary switch, where they cycle from an on and off state when GTP or GDP are bound, respectively. They are known to play a critical role in many cellular functions where a dysregulation could potentially lead to oncogenic behavior or other malignancies. In our laboratory, our focus is the study of a Ras related protein Cell division control 42 homolog (Cdc42) which belongs to the Rho subfamily. Cdc42 plays a critical role in many biological signaling processes; therefore, its uncontrol gene expression has been known to play a pivotal role in cancer cell progression and metastasis. A single point mutation was done on Cdc42 where a Threonine(T) was replaced to an Alanine(A) at position 35 (Cdc42(T35A)), this demonstrated a decrease in the flexibility in a region that is important for the interactions of downstream effector proteins.1 AZA197 has been demonstrated to show selectivity to Cdc42 and inhibit colon rectal tumor growth.2 The purpose of this research was to successfully synthesize the small molecule, AZA197, and study its interaction with Cdc42. Fluorescence spectroscopy was used in both characterizations of Cdc42(WT) and Cdc42(T35A) interactions with AZA197. The data suggest that in the presence of AZA197 Cdc42(WT) is more resistant to protein denaturation and has an increase in the Gibbs free energy of unfolding in the absence of GnHCl. Results also highlighted that AZA197 also interacts with Cdc42(T35A) and may have a destabilizing effect on this Cdc42 variant.

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