Undergraduate Honors Thesis Projects
Date of Award
Spring 2026
Document Type
Honors Paper
Degree Name
Biochemistry and Molecular Biology-BS
Department
Biochemistry and Molecular Biology
Advisor
Dr. John Tansey
First Committee Member
Dr. John Tansey
Second Committee Member
Dr. Brandon Sinn
Third Committee Member
Dr. Amy Johnson
Keywords
Lipid, metabolism, Perilipin 5, lipid droplet
Subject Categories
Biochemistry | Biochemistry, Biophysics, and Structural Biology | Higher Education | Molecular Biology
Abstract
Neutral lipids in animal cells are stored in structures called lipid storage droplets (LD). LDs are found in nearly all tissues of the body, consisting of neutral lipids forming a hydrophobic core, surrounded by a monolayer of phospholipids. Within the monolayer are proteins, including the perilipin family of proteins which regulate the storage and release of lipids. One family member, perilipin 5, works to coordinate lipid storage and breakdown while linking LDs to mitochondria to enhance energy production and metabolic regulation. Using AlphaFold, we obtained a predicted structure of perilipin 5, consisting of a boot-like shape containing a four-helix bundle and a hydrophobic pocket analogous to what has been observed experimentally in perilipin 3. In silico modeling data indicates that the hydrophobic pocket of perilipin 5 likely binds to hydrophobic molecules such as diacylglycerol. To probe the function of the hydrophobic pocket further, a mutant variant of perilipin 5 was created by replacing a key tryptophan in the hydrophobic pocket with glycine (W388G). The mutant was expressed in CHO cells and localized to lipid storage droplets when examined using immunofluorescence microscopy. Similar observations were made with cells expressing wild type protein indicating that the mutant protein is likely properly folded and functional and can be used to probe the nature of the pocket. To further probe the function of this pocket we have synthesized a fluorescent diacylglycerol analog and will be using it to validate our predictions made using in silico modeling.
Licensing Permission
Copyright, all rights reserved. Fair Use
Recommended Citation
Errington, Emma, "Structure Function Studies on the Hydrophobic Pocket of Perilipin 5" (2026). Undergraduate Honors Thesis Projects. 212.
https://digitalcommons.otterbein.edu/stu_honor/212
Acknowledgement 1
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Acknowledgement 2
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