Date of Last Revision
2023-05-02 18:58:49
Major
Chemistry - Biochemistry
Degree Name
Bachelor of Science
Date of Expected Graduation
Spring 2016
Abstract
This project investigates the cluster stability of an outer mitochondrial membrane protein mitoNEET under mutational stress and drug binding analysis. MitoNEET is a part of a class of proteins that coordinate an Fe-S molecule into its tertiary structure. This protein is believed to facilitate the oxidative capacity of the electron transport chain by reversible loss of its coordinated Fe-S molecule. The research of this paper uses site directed protein mutagenesis to selectively alter the three cysteine, one histidine cluster coordination into more common coordination patterns such as two cysteine, two histidine or four cysteine. The ultimate goal is to optimize conditions for future research and analysis by UV-Vis spectrometry. Research led to the conclusion that optimal conditions for kinetic analysis under acidic conditions were in a pH 5.5 20 mM citric acid reaction buffer with 100 mM NaCl and 5% DMSO. To the reaction buffer 100 mM protein and 100 mM drug were added. This reaction was carried out using the Shimadzu Spectra apparatus (Absorbance values at 459 nm) and the reaction cuvettes were temperature controlled by a water bath at 27°C to yield complete cluster loss within 6 hours.
Research Sponsor
Dr. Thomas Leeper
First Reader
Dr. Adam Smith
Second Reader
Dr. Claire Tessier
Recommended Citation
Mickley, Lisa, "Mutagenesis-Based Active-Site Characterization of the Diabetes Drug Target MitoNEET" (2016). Williams Honors College, Honors Research Projects. 278.
https://ideaexchange.uakron.edu/honors_research_projects/278
Included in
Analytical Chemistry Commons, Medicinal-Pharmaceutical Chemistry Commons, Other Chemistry Commons