Date of Graduation

Summer 2015

Document Type

Honors Research Project

Degree Name

Bachelor of Science

Major

Chemistry - Biochemistry

Research Sponsor

Dr. Adam Smith

First Reader

Dr. Leah Shriver

Second Reader

Dr. Sailaja Paruchuri

Abstract

Studies on the organization of membrane proteins such as GPCRs in live cells help to draw important conclusions regarding cellular communication, and have become a significant topic in drug therapy development. Observing proteins in MDCK cells, which are able to form three-dimensional cultures, provides an intermediate step to better understanding the organization and behavior of these proteins in a tissue-like culture. In this study, fluorescent stains were used on fixed cells to show the morphological differences between MDCK cells in monolayer cultures and three-dimensional cultures, which form hollow, spherical cysts of tightly packed cells. Monolayer cells were transfected to view the spatial organization of the GPCR protein, opsin, in live MDCK cells, and compared to the non-GPCR protein, plexin. Images from the transfected cells for both opsin and plexin show successful localization of the proteins to the cellular membrane in a relatively even dispersion, with no obvious sorting to any particular domain of the cell. This understanding of membrane protein organization in monolayer MDCK cultures provides a strong basis for future experiments to be performed on three-dimensional, tissue-like cultures of these cells.

Included in

Biochemistry Commons

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