RUI: The Role of Trehalose-Based Oligosaccharides and Glycomacromolecules on Protein Stability

RUI：海藻糖低聚糖和糖大分子对蛋白质稳定性的作用

• 批准号: 1708546
• 负责人: Nicole Snyder
• 金额: $ 29.4万
• 依托单位: Davidson College
• 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: 2017-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1708546&HistoricalAwards=false
• 关键词: RUI; Role; Trehalose; Based; Oligosaccharides

The Chemistry of Life Processes Program in the Chemistry Division funds this research. Professor Nicole L. Snyder of Davidson College investigates a unique carbohydrate, a disaccharide known as trehalose. This research provides a better understanding of the roles of trehalose and trehalose-based analogs in helping organisms survive environmental stresses such as extreme lack of moisture. The experiments use a combination of advanced synthetic and biochemical techniques to systematically study how the structure of trehalose and trehalose-based analogs influence their protective function. The undergraduate students supported by this work receive valuable training in advanced organic synthesis with an emphasis on carbohydrate chemistry, molecular spectroscopy, and biochemical and biophysical techniques that enhances their education and prepares them for a competitive workforce. In addition, this project supports the development of a new course on the emerging subject of Glycomics at Davidson College. An integral part of the project is also a dynamic outreach program that engages the wider community, including local teachers of the Charlotte Teachers Institute, in appreciating the broad role that carbohydrates play in life processes. The unique properties of trehalose and its ability to preserve a wide array of biomolecules under extreme stress (e.g. desiccation, thermal stress, and oxidation) have captured the attention of scientists for over one hundred years. Recently, several novel trehalose-based oligosaccharides (TOS) with unique structures and preservation properties have been identified, but their specific role in molecular preservation is unknown. The fundamental research in this work uses a multifaceted, systematic approach that combines chemical synthesis, molecular spectroscopy and biochemistry to synthesize and analyze a series of novel trehalose-based oligosaccharides (TOS) and trehalose-based precision glycomacromolecules (TPG) to determine how the unique structure of these compounds correlates to their function. Chemical synthesis is used to provide control over the structure of each compound. Once synthesized, each compound is analyzed by NMR spectroscopy to provide detailed structural information. A combination of biophysical studies, including saturation transfer difference (STD)-NMR, differential scanning calorimetry (DSC), and isothermal titration calorimetry (ITC), is used to study the solution and solid state associations between TOS/TPG analogs and several proteins selected based on their various stabilities towards desiccation. Finally, enzyme kinetics studies measure the function of enzymes stabilized with TOS/TPG to ensure secondary structure integrity as well as function after desiccation. The results from this research may provide a first look at the factors that govern the structure-function relationships of TOS/TPG and how these unique compounds help organisms survive extreme environmental stress.

化学系的生命过程化学项目资助了这项研究。戴维森学院的 Nicole L. Snyder 教授研究了一种独特的碳水化合物，一种称为海藻糖的二糖。这项研究使人们更好地了解海藻糖和海藻糖类似物的作用。帮助生物体在极端缺乏水分等环境压力下生存。实验结合了先进的合成和生化技术，系统地研究了海藻糖的结构和结构。基于海藻糖的类似物会影响其保护功能。这项工作支持的本科生接受了高级有机合成方面的宝贵培训，重点是碳水化合物化学、分子光谱以及生物化学和生物物理技术，以提高他们的教育水平并为他们成为有竞争力的劳动力做好准备。此外，该项目支持戴维森学院开发关于糖组学这一新兴学科的新课程，该项目的一个组成部分也是一个动态的外展计划，该计划吸引了更广泛的社区，包括夏洛特教师学院的当地教师。一百多年来，人们认识到碳水化合物在生命过程中发挥的广泛作用，以及它在极端压力（例如干燥、热压力和氧化）下保存多种生物分子的能力。最近，已经鉴定了几种具有独特结构和保存特性的新型海藻糖基寡糖（TOS），但它们在分子保存中的具体作用尚不清楚。多方面的系统方法，结合化学合成、分子光谱学和生物化学来合成和分析一系列新型海藻糖寡糖 (TOS) 和海藻糖精密糖大分子 (TPG)，以确定这些化合物的独特结构与其功能的相关性化学合成用于控制每种化合物的结构，合成后，通过核磁共振波谱分析每种化合物，以提供详细的结构信息。包括饱和转移差 (STD)-NMR、差示扫描量热法 (DSC) 和等温滴定量热法 (ITC)，用于研究 TOS/TPG 类似物与根据其不同稳定性而选择的几种蛋白质之间的溶液和固态关联。最后，酶动力学研究测量了用 TOS/TPG 稳定的酶的功能，以确保二级结构的完整性以及干燥后的功能。这项研究的结果可以提供对这些因素的初步了解。控制 TOS/TPG 的结构-功能关系，以及这些独特的化合物如何帮助生物体在极端环境压力下生存。