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

通过研究启动奖的历史黑人学院和大学本科课程(HBCU-UP)为历史黑人学院和大学的初级和职业生涯中期教师提供支持，这些教师正在建立新的研究项目或重新定向和重建现有的研究项目。预计该奖项有助于进一步提高教师的研究能力和有效性，并改善所在机构的研究和教学。这一奖项授予马里兰大学东部肖特分校，以支持教师、本科生和研究生寻求阐明糖基化对乳腺上皮细胞PGE2受体的功能和亚细胞定位的作用，并表征其细胞内糖基化特征。糖基化是最重要的蛋白质翻译后修饰之一，本项目旨在利用糖蛋白质组学、分子克隆、定点突变和免疫细胞化学等方法来促进对这一问题的认识。这个项目的主要焦点是PGE2，它是一种最丰富的生物活性脂质分子，参与了生理和病理生理过程，并通过与G蛋白偶联受体(GPCRs)结合来介导其作用。鉴于糖基化在细胞内分子机制中的重要性，本项目描述了以下工作：1)使用糖凝胶和糖蛋白组学技术分析乳腺上皮细胞中糖链的异质性；2)使用定点突变来确定糖基化对EP受体功能的影响；以及3)使用共聚焦显微镜分析糖基化对PGE2-EP1、EP1v和EP4受体亚细胞定位的影响。这个项目有可能促进对糖基化在调节细胞内钙和cAMP信号中的作用的了解，这两个信号是许多生物系统中的关键二级信使。此外，这些研究努力可能会扩大研究所的研究能力，并有助于未来的学生从细胞和分子机制上理解PGE2 EP1/EP4信号轴在乳腺细胞中的作用。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.