Date Written

4-2015

Document Type

Honors Paper

Degree Name

Biochemistry and Molecular Biology-BS

Department

Biochemistry and Molecular Biology

Advisor

John Tansey, Ph.D.

First Committee Member

John Tansey, Ph.D.

Second Committee Member

Jennifer Bennett, Ph.D.

Third Committee Member

Jessica Crossfield-McIntosh

Keywords

Perilipin 5, Immunofluorescence, Rab18, Rab32, PAT Family

Subject Categories

Biochemistry | Molecular Biology

Abstract

Perilipin 5 is one of the more recently discovered in the PAT protein family. Several studies on this protein have shown that it resides in multiple intracellular locations such as the endoplasmic reticulum, the cytosol, the mitochondria, and most interestingly on lipid droplets. These lipid droplets have proven not to be just clumps of fat, but to be very complex intracellular organelles, with a core of lipids surrounded by a phospholipid monolayer coated by specific proteins, one of those being perilipin 5. Some proteomic studies have revealed these lipid droplets to be involved in intracellular trafficking, signaling, cytoskeletal organization and lipid metabolism. By looking at other interaction partners of perilipin 5 we will have greater understanding of its function on the lipid droplet. In this experiment, I hypothesized that members of the Rab family of proteins should be considered as potential interacting partners, specifically Rab32 and Rab18. Immunofluorescence microscopy was performed on CHO cells under different antibody conditions and observed under various wavelengths of light. The images confirmed that perilipin 5 was located around lipid droplets, and that both Rab32 and Rab18 were expressing in the cells. When the Rab antibodies were combined with the perilipin 5 antibody, more colocalization was shown for Rab18 than for Rab32. There were also large amounts of endoplasmic reticulum fluorescence in many of the images. Collectively these data indicate that perilipin 5 and Rab 18 may work together to assist in droplet trafficking.

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