Novel reactivity of Fhit proteins: catalysts for fluorolysis of nucleoside 5′-phosphoramidates and nucleoside 5′-phosphosulfates to generate nucleoside 5′-phosphorofluoridates Available to Purchase

Fragile histidine triad (HIT) proteins (Fhits) occur in all eukaryotes but their function is largely unknown. Human Fhit is presumed to function as a tumour suppressor. Previously, we demonstrated that Fhits catalyse hydrolysis of not only dinucleoside triphosphates but also natural adenosine 5′-phosphoramidate (NH₂-pA) and adenosine 5′-phosphosulfate (SO₄-pA) as well as synthetic adenosine 5′-phosphorofluoridate (F-pA). In the present study, we describe an Fhit-catalysed displacement of the amino group of nucleoside 5′-phosphoramidates (NH₂-pNs) or the sulfate moiety of nucleoside 5′-phosphosulfates (SO₄-pNs) by fluoride anion. This results in transient accumulation of the corresponding nucleoside 5′-phosphorofluoridates (F-pNs). Substrate specificity and kinetic characterization of the fluorolytic reactions catalysed by the human Fhit and other examples of involvement of fluoride in the biochemistry of nucleotides are described. Among other HIT proteins, human histidine triad nucleotide-binding protein (Hint1) catalysed fluorolysis of NH₂-pA 20 times and human Hint2 40 times more slowly than human Fhit.