Engineering Factor Xa Protease Sites in the Exofacial Loop 9-10 of GLUT1

Cell membranes often allow small, uncharged solutes to pass freely, while larger molecular species such as nutrients and ions need the help of transporters. Glucose transporters (GLUT) are required for movement of glucose across the cell membrane. The structure and function of GLUT1 has been studied intensively for many years but much remains unknown. Previous research identified 12 transmembrane alpha helices, but a high resolution structure of GLUT1 is not available. The goal of this project was to insert a Factor Xa protease site in the exofacial loop 9-10 of GLUT1 to allow for cleavage of the protein at that loop for providing further insights into domains of the protein involved in the binding of substrates and inhibitors