Thermodynamical studies of oxygen equilibrium of hemoglobin. Nonuniform heats and entropy changes for the individual oxygenation steps and enthalpy-entropy compensation.

Precise oxygen equilibrium curves of human adult hemoglobin were determined by the automatic recording method at several temperatures in the presence and absence of 2,3-diphosphoglycerate (DPG) or inositol hexaphosphate (IHP) with 0.05 M 2,2-bis(hydroxymethyl)-2,2',2''-nitrolotriethanol (bis-tris) buffers (pH 7.4) containing 0.1 M Cl-. The equilibrium data were analyzed according to the Adair scheme, and the heats, deltaHi (i = 1,2,3,4) and the entropy changes, deltaSi (i = 1,2,3,4), for the individual oxygenation steps were obtained. The shape of the equilibrium curve varies on temperature changes whether DPG or IHP is present or absent. In consequence, the deltaHi value depends on i and on the presence of DPG and IHP. Behavior of deltaSi is similar to that of deltaHi. The similar behavior of deltaHi and deltaSi resulted in a compensation phenomenon. The contribution of T cdeltaSi to the free energy change is compensated by the contribution of deltaHi at the first three oxygenation steps but not at the fourth step, and for i = 1,2, and 3 changes of T cdeltaSi value upon the addition of DPG and IHP are compensated by accompanied changes of deltaHi value, where T c (= 260 K) is the compensation temperature. A major part of both the enthalpy-entropy compensation and nonuniformity of deltaHi and deltaSi appears to be attributable to contributions of the oxygen-linked binding of Cl-, DPG and IHP, by hemoglobin. The present results do not necessarily support the earlier idea of Wyman that the cooperative oxygenbinding is essentially an entropy effect.