Coupled reactions in hemoglobin. Heme-globin and dimer-dimer association.

Five different human hemoglobins were used to test the postulate that dissociation of hemoglobin (Hb) tetramers into alpha beta dimers and dissociation of heme from globin are linked reactions. Spectrophotometric measurements of the initial rate of heme transfer from Hb to serum albumin were made over a 3000-fold range of Hb concentration and yielded the heme-globin dissociation rate constant for tetramers and that for dimers. The tetramer-dimer dissociation constant (K4,2) could then be calculated from the rate constant at intermediate concentrations. The values obtained for the five hemoglobins, spanning a 250-fold range in K4,2, were in good agreement with those found by direct methods. The relation between this new linkage reaction of hemoglobin and the classical ones, such as the reciprocal relation between the binding of oxygen and protons, is discussed briefly.