Effective Mass Spectrometer (MS)-Based Methods for Systematic and Unambiguous Characterization of Protein O-GlcNAcylation

在化学系化学测量和成像项目的支持下，佐治亚理工学院的吴荣虎教授和他的团队正在努力提高我们表征一组重要的天然蛋白质修饰的能力，这些修饰涉及糖的附着。具体来说，吴博士正在开发基于质谱的生物技术工具，以更好地了解o - glcnac酰化，这是一种影响细胞功能的蛋白质修饰（例如基因表达的调节）。从事研究的研究生和本科生在仪器、化学测量和大规模数据分析方面接受了宝贵的培训。吴博士还为高中学生和教师提供研究机会，让他们接触到前沿研究，激发他们对STEM的兴趣。Wu小组试图通过开发新的质谱（MS）为基础的方法，全面而明确地表征蛋白质o - glcn酰化。这个问题是具有挑战性的，因为在复杂的生物样品中目标蛋白的天然丰度很低，并且需要区分含有O-GlcNAc的蛋白质和具有立体异构体O-GalNAc的蛋白质。吴博士正在开发选择性富集o - glcnac酰化蛋白的方案，使o - glcnac酰化蛋白能够进行全局和定量分析。细胞质和细胞核中糖蛋白的鉴定及其动力学的定量，将为我们理解蛋白o - glcn酰化提供有价值的信息，在生物学和生物医学研究中具有潜在的应用价值。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 support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Ronghu Wu and his group at Georgia Tech are working to improve our ability to characterize an important group of natural protein modifications that involve attachment of sugars. Specifically, Dr. Wu is developing biotechnology tools based on mass spectrometry, in search of better understanding of O-GlcNAcylation, a protein modification which impacts cell function (for example, regulation of gene expression). Graduate and undergraduate students engaged in the research receive valuable training in instrumentation, chemical measurements, and large-scale data analysis. Dr. Wu also provides research opportunities for high school students and teachers, exposing them to cutting-edge research and stimulating their interests in STEM. The Wu group seeks to comprehensively and unambiguously characterize protein O-GlcNAcylation through the development of novel mass spectrometry (MS)-based methods. The problem is challenging because of the low natural abundance of the target proteins in complex biological samples, and the need to distinguish proteins with O-GlcNAc from those with stereoisomeric O-GalNAc. Dr. Wu is developing protocols which selectively enrich O-GlcNAcylated proteins, enabling global and quantitative analysis of protein O-GlcNAcylation. Identification of glycoproteins in the cytosol and the nucleus, and quantification of their dynamics, will provide valuable information to facilitate our understanding of protein O-GlcNAcylation, with potential applications in biological and biomedical research.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.