Glycoproteins via Chemoselective Ligation of Proteins and Glycans

通过蛋白质和聚糖的化学选择性连接形成糖蛋白

• 批准号: 1665274
• 负责人: Philip Garner
• 金额: $ 46.5万
• 依托单位: Washington State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1665274&HistoricalAwards=false
• 关键词: Glycoproteins; via; Chemoselective; Ligation; Proteins

The Chemical Synthesis Program of the Chemistry Division supports this project by Professor Philip Garner. Professor Garner is a faculty member in the Department of Chemistry at Washington State University. He is developing new methodology for the synthesis of glycoproteins, which are found widely in eukaryotic cells and play important roles cell-cell signaling, immune response, and neuronal development. The study of glycoproteins has been limited by their inaccessibility. This project blends chemical and biological approaches to develop a new suite of tools for the conjugation of glycans to both unprotected peptides and ultimately proteins. Because of its interdisciplinary nature, it is well suited for the education of scientists at all levels. Professor Garner's group is also well positioned to provide the highest level of education and training for students underrepresented in science. Outreach activities involving local community college students are also part of the funded project.   The project develops methodology for the convergent synthesis of glycoproteins, specifically focusing on reactions that could be used for the assembly of glycophosphatidylinositol (GPI)-conjugated proteins and N-glycoproteins. Both of these posttranslational protein modifications are found widely in eukaryotic cells and play important roles in a variety of biological processes. The study of glycosylated proteins has been limited by the difficulty in obtaining these substances in homogeneous form. Currently available chemoenzymatic approaches either do not produce native structures (GPI-conjugated proteins) or are not general (N-glycoproteins). This project is developing a solution that is based on the Cu(II)-mediated coupling of protein thioacids and glycan amines. These Cu(II)-mediated couplings are distinguished from other ligation reactions by their enhanced rates and chemoselectivities, which appear to be a function of a unique reaction mechanism. The approach is novel and could, in theory, be applied to any GPI- and N-glycoprotein. GPI-proteins are expressed on the surface of neurons, fibroblasts, endothelial cells and many other cell types. The project trains both graduate and undergraduate students in organic synthesis and its application to biological molecules. This activity benefits our society by preparing a new generation of students for careers in either academia or the pharmaceutical/biotech sectors. The project addresses the currently limited interaction of students and faculty at local community colleges with scientists actively engaged in research at the university level through a planned symposium.

化学部化学合成计划支持菲利普·加纳教授的这一项目。加纳教授是华盛顿州立大学化学系的教员。他正在开发合成糖蛋白的新方法，糖蛋白广泛存在于真核细胞中，在细胞-细胞信号、免疫反应和神经元发育中发挥重要作用。糖蛋白的研究因其不可获得性而受到限制。该项目融合了化学和生物方法，开发了一套新的工具，用于将多糖连接到未受保护的多肽和最终的蛋白质上。由于它的跨学科性质，它非常适合于各级科学家的教育。加纳教授的团队也处于有利地位，可以为科学界代表性不足的学生提供最高水平的教育和培训。涉及当地社区大学生的外展活动也是该资助项目的一部分。该项目开发了糖蛋白聚合合成的方法，特别关注可用于组装糖磷脂酰肌醇(GPI)结合蛋白和N-糖蛋白的反应。这两种蛋白质翻译后修饰广泛存在于真核细胞中，并在多种生物学过程中发挥重要作用。糖基化蛋白质的研究由于难以获得均一形式的这些物质而受到限制。目前可用的化学酶方法要么不产生天然结构(GPI结合蛋白)，要么不是通用的(N-糖蛋白)。该项目正在开发一种基于铜(II)介导的蛋白质硫酸和葡聚糖胺偶联的解决方案。这些铜(II)介导的偶联反应与其他连接反应的不同之处在于它们的增强速率和化学选择性，这似乎是一种独特的反应机制的函数。这种方法是新颖的，理论上可以应用于任何GPI和N-糖蛋白。GPI-蛋白表达于神经元、成纤维细胞、内皮细胞等多种细胞表面。该项目培训研究生和本科生有机合成及其在生物分子中的应用。这项活动使我们的社会受益，为新一代学生在学术界或制药/生物技术部门的职业生涯做好准备。该项目解决了目前当地社区学院的学生和教职员工通过计划中的专题讨论会与积极参与大学一级研究的科学家之间有限的互动问题。

项目成果

Site-Specific Synthesis of Cysteine-Bridged Glycoproteins via Expressed Protein Glycoligation

通过表达蛋白糖基化位点特异性合成半胱氨酸桥糖蛋白

• DOI: 10.1021/acs.bioconjchem.0c00437
• 发表时间: 2020
• 期刊: Bioconjugate Chemistry
• 影响因子: 4.7
• 作者: Holloran, Nicholas;Collins, Daniel;Rathnayake, Upendra;Zhang, Bixia;Koh, Minseob;Kang, ChulHee;Garner, Philip
• 通讯作者: Garner, Philip