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

历史上的黑人学院和大学本科课程（HBCU-UP）通过研究启动奖为历史上的黑人学院和大学的初级和中级职业教师提供支持，他们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。预计该奖项将有助于进一步提高教师的研究能力和效率，并改善家庭机构的研究和教学。该奖项授予马里兰州东部肖特大学，支持教师，本科生和研究生的研究经验，因为他们试图阐明糖基化对乳腺上皮细胞中PGE 2受体的功能和亚细胞定位的作用，并表征其细胞内糖基化概况。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组学，分子克隆，定点突变和免疫细胞化学方法的组合来推进这方面的知识。该项目的主要重点是PGE 2，它是参与生理和病理生理过程的最丰富的生物活性脂质分子之一，并通过与G蛋白偶联受体（GPCR）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，本项目描述了以下努力：1）使用糖组学和糖蛋白质组学技术分析乳腺上皮细胞中的聚糖异质性; 2）使用定点突变来确定糖基化对EP受体功能的影响;（3）用共聚焦显微镜分析糖基化对PGE 2-EP 1、EP 1v和EP 4受体亚细胞定位的影响。该项目有可能推进糖基化在调节细胞内钙和cAMP信号传导中的作用的知识，这是许多生物系统中的关键二级信使。此外，这些研究工作可能会扩大该机构的研究能力，并有助于未来的学生对乳腺细胞中PGE 2 EP 1/EP 4信号轴的细胞和分子机制的理解。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。

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.