Phosphorylation of Anti-HIV Nucleoside Analogs by Nucleoside Diphosphate Kinase

AbstractThe reaction of NDP kinase with antiviral nucleoside triphosphates used in antiviral therapies was studied at the presteady state by fluorescence stopped-flow and compared with the steady-state parameters. The affinity of the analogs was determined by fluorescence titration of a mutated enzyme with an inserted Trp in the binding site. The lack of the 3′ hydroxyl in analogs is shown to decrease the kcatmore than the KD.