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. Author manuscript; available in PMC: 2014 Nov 4.
Published in final edited form as: Cell Rep. 2014 Mar 27;7(1):180–193. doi: 10.1016/j.celrep.2014.02.042

Figure 4. Heme Interacts with Tumor Suppressor p53 through Its C275AC277P Motif and Interferes with p53-DNA Interactions.

Figure 4

(A and B) Recombinant wild-type or p53C275,277A mutant protein (10 μM) was incubated with hemin (50 μM) on ice, followed by gel filtration and TMB assay (Pierce).

(C) A close-up view of the C275 and C277 residues of human p53 in the p53 core domain-DNA (Cho et al., 1994) (derived from PDB 1TSR).

(D) EMSA of the indicated hemin amounts, p53 protein, and double-stranded DNA probes containing p53RE consensus sequences. Band designations are free DNA (F) and bound DNA (B; p53-DNA complex).

(E) HCT116 p53−/− cells transiently expressing HA-tagged wild-type p53 were cotransfected with indicated luciferase reporter plasmids, then treated with or without hemin (10 μM, 6 hr). Error bars indicate ± SEM (n = 3). **p < 0.01.