Senior Honors Projects, 2020-current

Creative Commons License

Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Date of Graduation




Document Type


Degree Name

Bachelor of Science (BS)


Department of Biology


Christopher E. Berndsen

Ray Enke

Jon Monroe


Ubiquitin-fold modifier 1 (Ufm1) is a member of the Ubiquitin (Ub) family of proteins whose primary function is degradation of proteins through a sequential mechanism of chemical reactions. Though Ufm1’s specific roles are largely unknown, this family of proteins has shown to play a part in a wide variety of processes, including regulation of the cell cycle1, secretory functions of cells2,3, and blood clotting4. Ufm1’s mechanism of action proceeds with the aid of three enzymes: an E1, E2, and E3. Uba5 is the E1 activating enzyme that is specific to Ufm1, and its mechanism of action and active site chemistry is not well understood5,6. We do know that the enzyme contains a catalytic cysteine residue that is thought to be activated by another structure within the protein5–7. The goal of this study was to investigate potentially catalytic residues within Uba5 through modeling with YASARA8, generate mutant proteins with point mutations within these residues, and purify these mutants for structural modelling and activity assays, to compare to the wild type. Such residues were identified and selected for purification as part of this study. These proteins were expressed; however, problems with the expression and purification protocol prevented us from successfully purifying the mutations, and SAXS data showed that what we did purify contained significant aggregation. As future directions, we suggest various adjustments to the expression and purification method that we feel will lead to better yields, and therefore continuation of these aims.



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