SOLID-STATE 17O NMR STUDIES OF GLUCOSE AND DEVELOPMENT OF 17O NMR AS A NEW PROBE OF GLUCOSE METABOLISM IN HELA CANCER CELLS

Abstract

This thesis reports on synthesis of site-specifically 17O-labeled glucose and related molecules and their solid-state 17O NMR characterization. The thesis also describes the development of 17O NMR as a new probe of glucose metabolism in HeLa cancer cells. In particular, a total of seven 17O-labeled glucose compounds were made via chemical synthesis: [1-17O]-D-glucose, [2-17O]-D-glucose, [3-17O]-D-glucose, [4-17O]-D-glucose, [5-17O]-D-glucose, [6-17O]-D-glucose, and [6-17O]-2-deoxy-D-glucose. In addition, [6-17O]-D-glucose-6-phosphate and [1,1,5-17O3]-glutamine were prepared through enzymatic reactions. Solid-state 17O NMR experiments were performed to determine 17O quadrupole coupling and chemical shift tensors in all seven glucose compounds. This is the first time that any carbohydrate molecule has been completely characterized by solid-state 17O NMR. This thesis demonstrates that [6-17O]-D-glucose and [1,1,5-17O3]-glutamine can be used as new tracers for monitoring glucose/glutamine metabolism in live cells. Effects of insulin, oxygen concentration, and glutamine concentration on the glycolysis process in live HeLa cancer cells were monitored by 17O NMR in real time. Finally, some preliminary results of 17O NMR studies of hexokinase-glucose complex are reported.