Date of Award

5-3-2012

Degree Type

Thesis

Degree Name

Bachelor of Science (BS)

Department

Chemistry and Biochemistry

Abstract

Protein arginine methyltransferases (PRMTs) are involved In many major biological pathways in the human body. Processes that demonstrate arginine methylation by PRMTs include, but are not limited to, histone modification, DNA transcription, and post-translational protein modifications. Although recent research has allowed the identification of several PRMT isoforms and exposed their involvement in these processes, relatively little is known about the details of how these enzymes perform their biochemical duties. It is currently hypothesized that the N-terminus of PRMT variant I is involved in recognizing substrates and aiding in catalysis by virtue of a change in its conformation. To understand how the Nterminal tail (Nt) of human PRMTI may influence substrate recognition, we made a mutant of rat PRMTI where the N-terminus could hypothetically be switched from a " locked down" conformation into a "free" conformation using a simple small molecule. After sequence confirmation of the mutations, different expression parameters were tested to determine optimal expression. The mutant protein was then purified from the soluble portion of the expression cell growth using Ni affinity techniques. Activity was assessed in the presence and absence of the small molecule diothiothreitol (OTT). The unexpected results of the activity assay show that the lockdown mutant is insensitive to OTT. These results have caused us to consider several conjectural, yet fundamental, theories as the hypothesis of the project evolves and future work is carried out.

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Faculty Mentor

Dr. Joan M. Hevel