CAREER: Force spectroscopy enabled multivalent glycan-binding protein engineering

多糖是由多个糖单元组成的碳水化合物。它们是地球上最丰富的生物分子类型，在生物学中发挥着关键作用。细胞表面的多糖和蛋白质之间的相互作用可以改变这些细胞的功能和对疾病的反应方式。这个项目的重点是探索多糖是如何与蛋白质结合的，以及什么性质会影响它们的结合方式。将开发工具来快速测量多糖和蛋白质之间的相互作用，目的是了解如何更好地设计结合蛋白质。从这项工作中获得的见解将有助于开发改进的生物燃料和可以预防感染的更好的药物。此外，该项目将通过动手实验和向用户传授化学工程概念的智能手机应用程序，让小学生通过大学新生参与进来。该项目的目标是定义蛋白质与多糖结合的分子起源。这将有助于阐明蛋白质-多糖结合作用中结构和功能之间的关系。为了实现这一点，将开发声力谱(AFS)工具包。该工具包将能够同时检测多种糖蛋白相互作用。碳水化合物结合模块(CBM；A型)将用于开发和验证该技术。AFS将允许比传统技术更高的吞吐量，并提供对与生理相关的声力加载速率在几个数量级上变化的影响的探测。对于所有类型的CBM库，将阐明单分子和整体系综估计的结合参数之间的比较。最终，这项技术将被用来筛选改良的抗原和工业生物催化剂。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Glycans are carbohydrates composed of multiple sugar units. They are the most abundant type of biomolecule on Earth and play a critical role in biology. The interactions between glycans and proteins on the surface of cells can change how those cells function and how they respond to disease. The focus of this project is to explore how glycans bind to proteins and what properties affect how they bind. Tools will be developed to rapidly measure the interactions between glycans and proteins with the goal of understanding how to better design binding proteins. The insights gained from this work will aid in the development of improved biofuels and better drugs that can prevent infections. Additionally, this project will engage elementary students through college freshmen via hands on experiments and smart-phone apps that teach users chemical engineering concepts.The goal of this project is to define the molecular origins of protein binding to glycans. This will help to clarify the relationship between structure and function in protein-glycan binding interactions. To accomplish this, an acoustic force spectroscopy (AFS) toolkit will be developed. The toolkit will be capable of assaying multiple glycan-protein interactions simultaneously. Carbohydrate binding modules (CBM; type A) will be used to develop and validate the technique. AFS will allow for much higher throughput that traditional techniques and provide for probing the effect of physiologically-relevant acoustic force loading rate varied over several orders of magnitude. Comparisons between single-molecule and bulk-ensemble estimated binding parameters will be elucidated for a library of CBMs of all types. Ultimately, the technique will be utilized as a screen to identify improved antigens and industrial biocatalysts.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.