Date Written

4-2019

Document Type

Honors Paper

Degree Name

Biochemistry and Molecular Biology-BS

Department

Biochemistry and Molecular Biology

Advisor

Dr. John T Tansey

First Committee Member

Dr. Jennifer Bennett

Second Committee Member

Dr. Karen Steigman

Keywords

Perilipin 5, amino-terminus, structure, purification, homo-oligomerization, carboxyl-terminus

Subject Categories

Biochemistry | Structural Biology

Abstract

The United States of America, among many other countries around the world, is suffering from lipid-related disorders. Developing treatment options for these disorders that impact the lives of billions of individuals is an imperative task for scientists. In order to develop treatments and therapeutics, it is first necessary to understand the etiologies of these disorders and their molecular mechanisms. The perilipins are a heavily studied family of proteins found on the surface of lipid droplets that have an integral relevance in regulating lipolysis. Understanding the proteins involved in lipolysis can not only lead to a more complete understanding of the mechanisms of the lipid droplet catabolic process, but can extend knowledge into the pathologies whose etiologies are rooted in lipid metabolism. The progress of developing a deeper understanding of these molecules has been inhibited by the lack of structural data. To date, only one member of the perilipin family, perilipin 3, has structural data accompanying it. Even then, only a partial structure of the protein is known. Expanding the amount of structural data within this protein family would expedite and aid in determining further functions of the perilipins. The aim of this study is to expand the breadth of the structural information available for future studies. This project is aimed at determining the structure and characteristics of the amino-terminal region of the protein perilipin 5. A DNA construct designed to encode for the amino-terminal region of the perilipin 5 protein was designed. This protein fragment was then expressed in Escherichia coli and purified via metal affinity chromatography. The relative purity of the elution fractions was determined and the homo-oligomerization of perilipin 5 was explored utilizing these protein fragments. Finally, the optimization of this overexpression protocol was initiated.

Available for download on Tuesday, April 11, 2023

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