Submission Date

7-18-2024

Document Type

Paper- Restricted to Campus Access

Department

Biochemistry & Molecular Biology

Faculty Mentor

Amanda Reig

Comments

Presented during the 26th Annual Summer Fellows Symposium, July 19, 2024 at Ursinus College.

Project Description

G4DFsc is a de novo protein designed to mimic the metal-binding active sites of natural non-heme diiron enzymes. Through site-directed mutagenesis, specific amino acids can be targeted to affect conformational changes in the active site. In this study, H77D or H77E mutations were made in the E44H-G4DFsc plasmid to create an active site in which the histidine amino acids are trans to one another. Analysis of the mutated protein can further characterize the structure-function relationship of the protein’s hydrolase capabilities. The H77D and H77E mutagenesis was successful. Both proteins were produced and purified, allowing for future characterization of their biophysical properties, and DNA cleavage activity will be analyzed.

Restricted

Available to Ursinus community only.

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