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Representative examples of spectrophotometric titrations to determine iron affinities and extinction coefficients of the FeIII/protein complexes under conditions where Kd(Fe) < < [protein] (G1; Kd = 5 μM, an upper limit estimate), and Kd(Fe) > [protein] (S1; Kd = 200 μM). The gray line indicates the protein concentration at which the experiment was done. Lines were fit to a binding isotherm that takes the protein concentration into account (35).
