Research Initiation Award: Analysis of Glycoprotein Composition and Function of PGE2 EP Receptors in Mammary-derived Cells

University Of Maryland Eastern Shore, Princess Anne MD

Investigators

Abstract

The Historically Black Colleges and Universities Undergraduate Program (HBCU-UP) through Research Initiation Awards provide support for junior and mid-career faculty at Historically Black Colleges and Universities who are building new research programs or redirecting and rebuilding existing research programs. It is expected that the award helps to further the faculty member's research capability and effectiveness and improve research and teaching at the home institution. This award to the University of Maryland Eastern Short supports faculty, undergraduate and graduate student research experiences as they seek to elucidate the role of glycosylation on the function and subcellular localization of PGE2 receptors in mammary epithelial cells and characterize their intracellular glycosylation profile. Glycosylation is one of the most important protein post-translational modifications, and this project aims to advance knowledge to this end utilizing a combination of glycoproteomic, molecular cloning, site directed mutagenesis and immunocytochemistry approaches. The primary focus of this project is PGE2, one of the most abundant bio-active lipid molecules involved in both physiological and pathophysiological processes and mediates its effects by binding to G-protein-coupled-receptors (GPCRs). Given the importance of glycosylation in intracellular molecular mechanisms, this project describes efforts to 1) analyze the glycan heterogeneity in mammary epithelial cells using glycomic and glycoproteomic techniques; 2) use site-directed mutagenesis to determine the effect of glycosylation on EP receptor function; and 3) use confocal microscopy to analyze the effect of glycosylation on subcellular localization of PGE2-EP1, EP1v and EP4 receptors. This project has the potential to advance knowledge on the role of glycosylation in regulating intracellular calcium and cAMP signaling, which are key secondary messengers in many biological systems. In addition, these research endeavors are likely to expand the research capacity of the institution and contribute to future students on the cellular and molecular mechanistic understanding of the PGE2 EP1/EP4 signaling axis both in mammary cells. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.