A Novel Platform for the Tunable Production of Proteoglycans

在大多数哺乳动物细胞的表面都是带有氨基和糖基（糖胺聚糖；GAGs）作为侧链连接的蛋白质。这些蛋白质被称为蛋白聚糖，对人体健康很重要。这些蛋白质的结构如何影响它们的功能仍然知之甚少。我们将开发一种可以调谐产生不同GAG的烟草细胞，以便更清楚地了解链长度，GAG组成和核心蛋白功能之间的关系。控制具有确定结构的gag的生产的能力将增强其在治疗应用中的潜在用途。本科生和研究生将接受培训，来自代表性不足群体的学生将被积极招募到实验室进行研究。学生将获得专业知识，这将更好地为他们在不断扩大的生物制造劳动力的生产参与做好准备。该项目的总体目标是证明在没有硫酸或其他修饰的情况下烟草中核心蛋白聚糖结构的生产。为了实现这一目标，我们需要扩展植物合成生物学的分子工具箱，使对蛋白质表达的额外控制成为可能。长期目标是在N. benthamiana中开发一个蛋白多糖生产平台，使我们能够生产具有确定的gag，链长，组成和核心蛋白的蛋白多糖。这种模块化的蛋白聚糖生产方法将使未来在各种生物环境中发现蛋白聚糖的结构-功能关系。反过来，这将允许开发一类新的功能性生物大分子，促进糖组学领域的革命。这项新技术将使研究人员能够探索gag和蛋白多糖的巨大潜力，以开发新的生物和医疗技术。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。

On the surface of most all mammalian cells are proteins that have amino and sugar groups (glycosaminoglycan; GAGs) attached as side chains. These proteins, known as proteoglycans, are important to human health. How the structure of these proteins influences their function is still poorly understood. We will develop a tobacco cell that can be tuned to produced different GAGs so the relationships between chain length, GAG composition, and core protein on function can be understood more clearly. The ability to control the production of GAGs with defined structure will enhanced their potential use in therapeutic applications. Undergraduate and graduate students will be trained, and students from underrepresented groups will be actively recruited to perform research in the labs. Students will gain expertise that will better prepare them for productive participation in the expanding biomanufacturing workforce.The overall objective of this project is to demonstrate the production of the core proteoglycan structure in tobacco without sulfation or other modifications. To accomplish this, we will need to expand the molecular tool box for plant synthetic biology, enabling additional control over protein expression. The long-term goal is to develop a proteoglycan production platform in N. benthamiana that will enable us to produce proteoglycans with defined GAGs, chain length, composition and core proteins. This modular approach to proteoglycan production will enable future discoveries into the structure-function relationship of proteoglycans in a variety of biological contexts. Which in turn will allow for the development of a new class of functional biological macromolecules, fostering a revolution in the field of glycomics. This novel technology will enable researchers to explore the enormous, untapped potential of GAGs and proteoglycans for the development of new biological and medical technologies.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.