Date of Award
2017
Document Type
Open Access Master's Thesis
Degree Name
Master of Science in Chemistry (MS)
Administrative Home Department
Department of Chemistry
Advisor 1
Marina Tanasova
Committee Member 1
Shiyue Fang
Committee Member 2
Tarun Dam
Abstract
Carbohydrate transporters or GLUTs of the major facilitator superfamily (MFS) are responsible for transporting sugars into the cell and have been of research interest for decades. Disruptions, mutations, and over-activations of GLUTs have been linked to a number of major diseases including cancer, obesity, and diabetes. Differentiating between transporters is incredibly difficult due to highly conserved structures, and so specific targeting between transporters has proven a complex challenge. GLUTs are highly flexible in their conformations however exactly what will and will not pass through the transporter is ambiguous at best, and many attempt to target these transporters have failed.
In an attempt to further understand GLUT5’s transport capacity and specificity several probes were created by conjugating 1-amino-2,5-anhydro-D-mannitol with a number of fluorescent coumarins. These probes were then tested in cancer and normal breast cell lines to determine uptake mechanisms and transport specificity. To determine transport specificity probes were tested in the presence of competitive and non-competitive inhibitors. Probe analysis was carried out by evaluating the gained fluorescence of treated cells in a microplate setting and through confocal microscopy. Confocal imaging and Z-stack was utilized to understand the ability of the probe to pass into the cytosol or to remain in the cellular membrane. As a result, probes reflecting uptake capacity vs. membrane expression of the transporter were developed. The cumulative analysis of structure-uptake relationship for the developed probes gives insight into the capability of GLUT5 cargo transport and as well as a method for imaging GLUT5 in the cellular membrane.
Recommended Citation
Fedie, Joseph, "Fluorescent Probe Development for Fructose Specific Transporters in Cancer", Open Access Master's Thesis, Michigan Technological University, 2017.