Spectrophotometrical Assay: FBPase activity was measured spectrophotometrically by employing the coupling enzymes phosphoglucose isomerase and glucose-6-phosphate dehydrogenase.17 The reduction of NADP+ to NADPH was monitored directly at 340 nm. Specifically, buffer (0.2 M Tris, 4 mM MgCl2, 4 mM (NH4)2SO4, 0.1 EDTA, pH 7.5), 0.2 mM of NADP+, 1.4 units of phosphoglucose isomerase and 0.5 units of glucose-6-phosphate dehydrogenase, 0-300M of inhibitor and 9 ng of FBPase, were mixed in a cuvette and equilibrated at 30° C. 70M of FBP was then added to initiate the reaction. Absdata were collected as a function of time using a JASCO V-630 spectrophotometer.
Spectrophotometrical Assay: FBPase activity was measured spectrophotometrically by employing the coupling enzymes phosphoglucose isomerase and glucose-6-phosphate dehydrogenase.17 The reduction of NADP+ to NADPH was monitored directly at 340 nm. Specifically, buffer (0.2 M Tris, 4 mM MgCl2, 4 mM (NH4)2SO4, 0.1 EDTA, pH 7.5), 0.2 mM of NADP+, 1.4 units of phosphoglucose isomerase and 0.5 units of glucose-6-phosphate dehydrogenase, 0-300M of inhibitor and 9 ng of FBPase, were mixed in a cuvette and equilibrated at 30° C. 70M of FBP was then added to initiate the reaction. Absdata were collected as a function of time using a JASCO V-630 spectrophotometer.