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Binding of mutant HemRs and wild-type HemR (HemRWT) to heme- and Hb-agarose. Lanes: M, molecular size markers; 1, OMs of E. coli expressing HemRWT; 2, Hb-agarose-bound HemRWT; 3, heme-agarose-bound HemRWT; 4, Hb-agarose-bound HemRH128K; 5, heme-agarose-bound HemRH128K; 6, Hb-agarose-bound HemRH461L; 7, heme-agarose-bound HemRH461L. Differences in the amount of material bound to heme and Hb-agarose are due to the 30-fold-higher binding capacity of the heme- over Hb-agarose.
