CAREER: Illuminating O-GlcNAc-Driven Functions of the Human Proteome

在化学系生命过程化学项目的支持下，来自韦恩州立大学的Charlie Fehl正在开发一种工具，这种工具可以特异性地、快速地传递，并且可以获得关于糖修饰对任何感兴趣的蛋白质的影响的信息。目前，已知人类细胞中三分之一的蛋白质在食用碳水化合物后会暂时被糖包裹，但这些糖的功能仅占所有人类蛋白质的3%左右。如果有能力确定高糖对细胞蛋白质的影响，阐明这些蛋白质的功能是可能的。这一新知识对于理解人体新陈代谢和我们的新陈代谢是如何被调节的非常重要。在这项研究中开发的化学工具可以使糖生物学的研究取得进展。Fehl博士的工作将侧重于培养多个层次的学生，从研究生和本科生，他们将进行研究，以及高中生，他们的参与项目将使他们意识到并了解STEM职业道路。一个关键的教育发展是“数据的演讲暴露/分析”（LEAD）模块，该模块在课堂上使用拟议实验中产生的数据训练本科生的假设形成和假设检验技能。该研究项目旨在利用“CRISPR”基因标记系统建立对高糖和低糖水平做出反应的蛋白质特异性分析。具体来说，韦恩州立大学的研究小组将操纵O-GlcNAc糖（o -连接n -乙酰氨基葡萄糖），这是所有人类细胞中主要的葡萄糖和营养传感分子。这三种建议的检测方法有望分别揭示高O-GlcNAc和低O-GlcNAc对这些检测中标记的特定蛋白的稳定性、亚细胞位置和整体细胞功能。稳定性分析将集中于一组o - glcn酰化激酶，这是细胞信号传导的主要驱动因素，可能对营养水平作出反应。定位分析将集中于一组已知的o - glcn酰化转录因子，这些转录因子参与调节细胞功能。“糖标签”系统有望提高细胞中特定蛋白质上的O-GlcNAc水平，使测量这些糖修饰对细胞代谢、信号传导和生存的影响成为可能。这三种新检测的结果将被汇编成一个功能数据库，与公众和科学界共享。该项目产生的数据有望迅速推进对糖驱动蛋白功能的了解，并为人体组织中代谢活动和体内平衡的调节提供新的见解。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

With the support of the Chemistry of Life Processes Program in the Division of Chemistry, Charlie Fehl from Wayne State University is developing tools that can be delivered specifically and rapidly and can elicit information on the effect of sugar modifications on any protein of interest. Currently, it is known that one third of the proteins in human cells are temporarily coated in sugars after eating carbohydrates, but the functions of those sugars are known for only about 3% of all human proteins. The elucidation of the functions of these proteins could be possible if one has the ability to determine the effects of high sugar on cellular proteins. This new knowledge is highly significant for the understanding of human metabolism and how our metabolism is regulated. The chemical tools being developed in this research could enable advancements in the study of sugar biology. Dr. Fehl’s work will focus on training students at multiple levels, from graduate and undergraduate students who will carry out the research as well as high school students whose participation in the project will enable them to become aware and informed about STEM career pathways. A key educational development is a “Lecture Exposure/Analysis of Data” (LEAD) modules that train undergraduates in hypothesis formation and hypothesis testing skills in the classroom using the data generated in the proposed experiments.This research project aims to build protein-specific assays that respond to high and low sugar levels using the “CRISPR” gene tagging system. Specifically, the Wayne State team will manipulate the sugar O-GlcNAc (O-linked N-acetylglucosamine), the major glucose- and nutrient-sensing molecule in all human cells. The three proposed assays are expected to reveal the stability, subcellular location, and overall cellular functions, respectively, of high vs. low O-GlcNAc on specific proteins tagged in these assays. Stability assays will focus on a set of O-GlcNAcylated kinases, which are major drivers of cell signaling that potentially responds to nutrient levels. Location assays will focus on a set known O-GlcNAcylated transcription factors, which are involved in regulating cellular functions. The “GlycoTag” system is expected to elevate O-GlcNAc levels on specific proteins in cells, making possible the measurement of the effects of these sugar modifications on cell metabolism, signaling, and survival. The results of these three new assays will be compiled into a functional database shared with the public and the scientific community. The data generated in this project are expected to rapidly advance knowledge of sugar-driven protein functions and provide new insights into how metabolic activities and homeostasis are regulated in human tissues.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.