A CONVENIENT METHOD FOR ENZYMATIC-SYNTHESIS OF RADIOLABELED GLUCOSE-1,6-BISPHOSPHATE

A convenient method to synthesize radiolabelled glucose-1,6-bisphosphate (Glc-1,6-P-2) of high specific activity and in a high yield is reported. The method is based on enzymatic formation of glucose-6-P from glucose and ATP using hexokinase. The labelled glucose-6-P is then converted to Glc-1,6-P-2 through an equilibrium reaction catalyzed by phosphoglucomutase. The two enzymatic steps and TLC separations necessary to afford radiochemically pure Glc-1,6-P-2 can be completed in one day. The outlined experimental procedure details the synthesis of [P-33]Glc-1,6-P-2 evenly labelled at the carbon 1 and the carbon 6 position. However the commercial availability of gamma-P-33 and gamma-P-32 labelled ATP and of glucose labelled with H-3, C-14 and C-13 provides the option to label glucose-1,6-bisphosphate with any of these isotopes.