X-ray Crystallography and Biochemical Analysis of Fumarase C Variant H129N
JSHS · 2020
Overview
University of Wisconsin-La Crosse Crystallographic studies of the structures of proteins help explain how those proteins function as enzymes. The field of crystallography has advanced in the past decade, mainly in terms of the energy that can be achieved by a beam in X-ray crystallography. Ho wever, current crystallography data on the structure of the H129N mutated variant of the enzyme fumarase has a significantly lower resolution than modern structures of the regular wild type. In this study, we used purification techniques such as Ni2+ metal-chelate chromatography to create crystals of the H129N variant with hanging drop vapor diffusion so we could solve its structure with X-ray crystallography, eventually obtaining a resolution of approximately 1.41 angstroms, near that of modern structures for the wild type. We also began experiments with circular dichroism and Michaelis-Menten kinetics to determine thermodynamic quantities for folding in wild type fumarase Cand the H129N variant as well as kinetic quantities for the catalysis of both enzymes in the dehydration of S -malate to fumarate, with the goal of understanding how the solved structure affects the function of the variant.
Awards (1)
- Poster Peer Awardee
Competition history
- JSHS 2020
Resources
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