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2 μM apo-DddW (under tight-binding conditions) was titrated with increasing concentrations of Fe(NH4)2(SO4)2 and the fluorescence intensity was monitored. The titration data were analyzed by nonlinear curve fitting using Eq (3) to produce the solid line. Upon data fitting, the stoichiometric ratio of Fe(II) to DddW monomer was determined to be 1:1.
