Date of Award

Spring 5-4-2025

Document Type

Distinction Paper

Degree Name

Biochemistry and Molecular Biology-BS

Department

Biochemistry and Molecular Biology

Advisor

John Tansey

First Committee Member

John Tansey

Second Committee Member

Carrigan Hayes

Third Committee Member

Joy Shoemaker

Keywords

Perilipin 5, Click Chemistry, Lipid Droplets, IR Dye, Phosphoethanolamine, Diacylglycerol

Subject Categories

Chemistry | Higher Education | Organic Chemistry

Abstract

Neutral lipids in mammalian cells are found in lipid storage droplets. Perilipin 5 is a lipid storage droplet protein, expressed predominantly in tissues with high oxidative metabolic activity, including the heart, muscle, and liver. Perilipin 5 plays a critical role in metabolic processes such as lipolysis, fatty acid storage, lipid droplet formation, and transport of lipids to mitochondria, therefore it is essential in maintaining metabolic homeostasis. AlphaFold predicts the structure of Perilipin 5 to have a conserved hydrophobic pocket of unknown function. Prior work has predicted that this pocket binds to diacylglycerol, suggesting a role in lipid metabolism. To better understand the structure-function relationship of this pocket, we designed a fluorescent diacylglycerol analog to use as a probe. The analog was synthesized through a click coupling reaction between 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and the N-hydroxysuccinamide ester of IRDye 680RD from LiCor Biotechnology. The synthesis was confirmed via thin-layer chromatography (TLC) under neutral, and acidic mobile phase conditions. To test these probes’ usefulness, we separated plasma lipoproteins by agarose gel electrophoresis, stained them with the probe, and visualized them. Discrete bands corresponding to each major class of lipoproteins were seen, indicating that this probe could be used to stain particles of bulk lipids. Treatment of samples with detergent (Tween 20, 10%) or Candida reugosa lipase (1 Unit, 10 minutes) altered banding patterns indicating that the structures had been disrupted. These data indicates that we have successfully synthesized a near-IR lipid probe which can be used to further study these systems.

Licensing Permission

Copyright, all rights reserved. Fair Use

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Acknowledgement 2

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