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Kinetics of heme binding to GAPDH. Heme binding to GAPDH was assessed by stopped-flow. The reaction proceeded cleanly to form ferric GAPDH-heme in a single step process (Panel A). The initial and final spectra given in the inset were collected at 0.1 s and 40 s, respectively. Plot of the observed rate constants for different doses of GAPDH yielded a binding constant kon of 17,800 M−1s−1 at 10 °C (Panel B).
