Bioinformatic Analysis and Design of a GLUT3 Construct to Examine Cell Surface Expression

Abstract

Glucose transporter type 3 (GLUT3) expression at cell surface plays a vital homeostatic function in neurons. However, the mechanism involved has yet to be fully examined. We devise a strategy to tag a rat GLUT3 cDNA clone. This will be used to assess GLUT3 localization at cell surface. To install the tag, we attempt to generate a point mutation at the first exofacial loop, which would result in a unique Hpa1 restriction site in GLUT3. In another strategy, to compare total versus surface GLUT3, we propose an HA-GLUT3-eGFP fusion protein. Bioinformatic analyses suggest that this protein would be functional. GLUT3 movement to plasma membrane is carried out through vesicles. These vesicles interact with many trafficking proteins that regulate GLUT3 surface expression. Data mining using the STRING database has identified potential GLUT3-interacting proteins. Future work will focus on expressing tagged-GLUT3 and examining interactions with these proteins.