The Expression of 19F-Labeled Beta-Phosphoglucomutase and the Evaluation of Its Inhibitors

Abstract

β-Phosphoglucomutase (βPGM) is an isomerase that catalyzes the conversion of β-glucose 1-phosphate to glucose 6-phosphate via a 1,6-bisphosphate intermediate. Incorporation of 5-fluorotryptophan (5FW) into βPGM (ie. 5FWβPGM) as a 19F NMR spectroscopic probe revealed that the 19F nucleus in 5FW is a sensitive probe for monitoring metal fluoride transition state analogue (TSA) complexation and ligand binding. The ability of βPGM to form transition state analogue complexes with novel compounds, β-glucose 1,6C-phosphonate, and β-2-deoxy-2-fluoro glucose and mannose 1C-phosphonates were examined; however, only β-glucose 1,6C-phosphonate (IC50 = 186 +/- 73 µM) and β-2-deoxy-2-fluoro glucose (2.68 +/- 0.04 µM) configured analogs bound to βPGM. All compounds that exhibited complexation with 5FWβPGM by 19F NMR spectroscopy were confirmed as competitive. Approaches to the synthesis of 1,6-bisphosphate and phosphofluoridates analogs as mechanism-based inactivators of 5FWβPGM are described.