CURF Introduction – Jordan Kuwik

My project, directed under the guidance of Dr. Kabirul Islam, focuses on the alpha-ketogluterate-dependent hydroxlyse (AlkB) class of proteins. These proteins demethylate a variety of different mRNA, tRNA, ssDNA, and dsDNA constructs. In doing so, they play a role in the stability of proteins and DNA/RNA fidelity within the cell. More interestingly, recent research has correlated mutants of these proteins to the development of genetically predisposed obesity and cancer in individuals. My project focuses on understanding the structural activity of these proteins that allows them to perform their demethylation activity. Specifically, each of these enzymes is activated by a small molecule activator, alpha-ketogluterate (2KG). 2KG is a natural intermediate of the TCA cycle and is thus found freely in mammalian and some bacterial organisms. This class of proteins is correspondingly inhibited by a structural analog of 2KG, N-oxylylglycine (NOG). NOG is not found naturally in mammalian cells, and can thus be used as a general inhibitor of these proteins in vitro. In order to understand the structural activity of these proteins, I perform site-directed mutagenesis to key amino acids (AAs) in the active site of each protein, as determined via crystallography. Specifically, I mutate larger AAs to those of smaller AAs (i.e. tyrosine to glycine), thus equivocally inserting a ‘hole’ into the active site. These mutated proteins are still activated by native 2KG but are no longer inhibited by WT NOG. With these mutant proteins, I then synthesize (with the help of my graduate mentor, Valerie Scott) NOGs with ‘bumps’ on them (i.e. chemical side chains that can fit into the ‘hole’ created after mutating the active site of the WT proteins). My principal hypothesis is that mutant proteins with altered active site AAs that are no longer inhibited by WT NOG will be inhibited by these bumped NOGs. This is because the ‘hole’ will effectively be filled by the ‘bump’ of the NOG. Due to the aforementioned correlation between mutants of these proteins predisposing individuals to genetically related obesity and cancer, understanding the mechanisms behind these proteins will more effectively bring forth the development of therapeutics to affected individuals.

My career goal is to become a physician. However, more than just focusing on the clinical aspect of medicine, I would like to conduct research focused on the biochemical and molecular aspects of medicine, much like my current research. I believe the CURF will help me pursue these goals of mine by allowing me to delve deeper into my current research and further experience the true nature of research. In doing so, the lessons of teamwork, trial-and-error, and a general understanding of how research is conducted will to my future of becoming a physician-scientist.

I am Microbiology major with minors in Chemistry, History, and Neuroscience, along with a Conceptual Foundations of Medicine Certificate. One unique thing about me is that I have been to all fifty states, with the last of the fifty being Florida!

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