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，来自州立大学的韦恩的Charlie Fehl正在开发工具，这些工具可以特异性地快速交付，并可以引出有关糖修饰对任何感兴趣的蛋白质的影响的信息。目前，已知人类细胞中有三分之一的蛋白质在食用碳水化合物后会暂时被糖包裹，但这些糖的功能仅为所有人类蛋白质的约3%所知。如果人们有能力确定高糖对细胞蛋白质的影响，就有可能阐明这些蛋白质的功能。这一新知识对于了解人体代谢以及我们的代谢是如何调节的具有重要意义。这项研究中开发的化学工具可以促进糖生物学研究的进展。费尔的工作将侧重于培训学生在多个层次，从研究生和本科生谁将进行研究，以及高中学生的参与该项目将使他们成为了解和了解干的职业道路。其中一个重要的教育发展是“讲座曝光/数据分析”（LEAD）模块，该模块利用拟议实验中产生的数据，在课堂上培养大学生的假设形成和假设检验技能。本研究项目旨在利用“CRISPR”基因标签系统构建对高糖和低糖水平做出反应的蛋白质特异性测定。具体来说，韦恩州立大学的研究小组将操纵糖O-GlcNAc（O-连接的N-乙酰葡萄糖胺），这是所有人类细胞中主要的葡萄糖和营养物质传感分子。预期三种拟定测定法分别揭示这些测定法中标记的特定蛋白质上的高与低O-GlcNAc的稳定性、亚细胞位置和总体细胞功能。稳定性试验将重点关注一组O-GlcNAc酰化激酶，这些激酶是细胞信号传导的主要驱动因素，可能对营养水平产生反应。定位分析将集中在一组已知的O-GlcNAc酰化的转录因子，这是参与调节细胞功能。“GlycoTag”系统有望提高细胞中特定蛋白质的O-GlcNAc水平，从而可以测量这些糖修饰对细胞代谢、信号传导和存活的影响。这三项新检测的结果将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糖驱动蛋白质功能的知识，并为人体组织中代谢活动和体内平衡如何调节提供新的见解。该奖项反映了NSF的法定使命，并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，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.