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Measurement of Zn2+ binding to MntC via a PAR competition assay. A binding isotherm was generated from 200 μM PAR and 50 μM Zn2+ titration with increasing concentration of MntC (0 to 80 μM). Inset, representative spectra obtained during MntC titration. The downward arrow denotes the decrease in absorbance at 500 nm, indicating the removal of Zn2+ from the PAR2Zn complex by adding (upward arrow) various concentration of MntC to form MntC-Zn2+.
