19-F NMR studies of the binding of a  fluorine-labeled phosphonate ion to E. coli alkaline phosphatase.
The interaction of a fluorinated phosphonate with Zn-2+-and Mn-2+-alkaline phosphatase as studied by 19-F NMR revealed a stoichiometry of 1:1 for the binding of the phosphonate anion to the enzyme. In the presence of two metal ions, one fluorinated phosphonate ion was found to interact strongly with the enzyme, while a different interaction was observed when the number of metal ions per enzyme exceeded two. Phosphate replaced enzyme bound phosphonate, as is shown by the 19-F NMR spectra. No direct interaction between the fluorinated phosphonate and the metal ion responsible for enzyme activity was indicated by the 19-F NMR data. This observation supports the idea of a considerable distance between metal ion and substrate binding site in Escherichia coli alkaline phosphatase.
D000469:Alkaline Phosphatase / Q000032:analysis*; D001665:Binding Sites; D004926:Escherichia coli / Q000201:enzymology*; D005461:Fluorine; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008345:Manganese; D063065:Organophosphonates; D011485:Protein Binding; D015032:Zinc
D063065:Organophosphonates; D005461:Fluorine; D008345:Manganese; D000469:Alkaline Phosphatase; D015032:Zinc