CAREER: Force spectroscopy enabled multivalent glycan-binding protein engineering

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.