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Histidine 32 and arginine 214 are individually important for GtrR binding to heme in vitro. Absorption spectra of increasing concentrations of heme (5–50 µM) in the presence of 5 µM purified GtrRH32A (A) or GtrRR214A (B). The spectrum corresponding to 50 µM free heme is shown as a solid pink line. Heme concentrations ranging from 5 to 50 µM of heme are graphed in grayscale from light to dark.
