Date of Graduation

Spring 2015

Document Type

Honors Research Project

Degree Name

Bachelor of Science

Major

Chemistry - Biochemistry

Research Sponsor

Dr. Thomas Leeper

First Reader

Dr. Sara Carlson

Second Reader

Dr. Claire Tessier

Abstract

MitoNEET, a mitochondrial membrane protein with an unsymmetrical Fe-S cluster coordination, was compared to similar proteins within the Rieske and Ferredoxin families. Point mutations were introduced via site-directed mutagenesis to induce stability, greater retention of the cluster, and to uncover structural limitations of the protein. Mutations were made within the cluster coordination and within the hydrophobic core of the protein. Some of the cluster mutations displayed a natural affinity for nickel during immobilized metal-ion affinity chromatography. It is possible that these mutations aid in the formation of a chelate when exposed to nickel. Spectroscopic monitoring of changes in cluster stability under low pH conditions was used to compare cluster retention across mutations. Some of the cluster mutants displayed a significant increase in stability versus wild type and other mutated forms of the protein. The hydrophobic mutations were found to push structural limitations of the protein by disrupting the hydrophobic core, even in the presence of the stabilizing cluster mutations. These mutation results would benefit from further structural research.

Included in

Biochemistry Commons

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