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GAPDH binds intracellular heme in mammalian cells, and this binding is disrupted by the H53A mutation. A, representative HPLC fractions with 14C counts from HA pulldowns. HA–GAPDH WT 14C peak corresponds to heme elution. B, 14C counts from HA pulldowns of HA–GAPDH-transfected GlyA cells. Values represent mean ± S.E. n = 4. Western blots of GlyA cells show expression of HA–GAPDH and β-actin. *, p ≤ 0.02; **, p < 0.005; ***, p < 0.0001, two-tailed t tests. mAU, milli-absorbance units.
