Oxygen binding constants for human hemoglobin tetramers.
High-precision studies of oxygen binding in hemoglobin (HbA0) solutions at near-physiological concentrations (2-12 mM heme; pHs 7.0-9.1; various buffers) have led to an unanticipated result: an unmeasurably low contribution from the triply ligated species. We have obtained this result from new differential oxygen-binding measurements for human hemoglobin through the use of a thin-layer apparatus, which enables study of solutions at high Hb concentrations. The effect of tetramer dissociation into dimers, which becomes significant at hemoglobin concentrations below 1 mM in heme, is avoided. The analysis of the binding reactions is thus cast in terms of tetramer-binding polynomial written with overall Adair equilibrium constants which directly reflect the contributions of intermediate ligated species. The unmeasurable contribution of the triply ligated species renders the equilibrium constants of the third and fourth stepwise reactions practically undeterminable.
D001665:Binding Sites; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008722:Methods; D010100:Oxygen / Q000378:metabolism*; D011312:Pressure; D011487:Protein Conformation
D006454:Hemoglobins; D010100:Oxygen