Submission Date

7-20-2018

Document Type

Paper- Restricted to Campus Access

Department

Biochemistry & Molecular Biology

Second Department

Chemistry

Faculty Mentor

Amanda Reig

Student Contributor

Alana Huynh

Comments

Presented during the 20th Annual Summer Fellows Symposium, July 20, 2018 at Ursinus College.

Supported by a National Institutes of Health Academic Research Enhancement Award (AREA) grant (R15-GM110657).

Project Description

The unique structure-function relationship of proteins has made de novo metalloprotein models a valuable tool in obtaining a deeper understanding of these essential biological molecules. To study the impact of active site configuration on reactivity, we use a de novo four helix bundle protein (DFsc) containing a non-heme di-iron center in which the active site has been varied. These variants were overexpressed in E. coli cells and purified via HPLC. The purified protein variants were then characterized through differing methods: (1) chromogenic assays to measure the reactivity of the proteins, (2) cyclic voltammetry to investigate their electrochemical properties, and (3) X-ray crystallography to determine the specific structure of the variants. This understanding of the protein structure will provide insight on how the different active site variations affect midpoint potentials and ultimately its reactivity.

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