Kinetic studies on the cupric ion oxidation of sheep hemoglobin.

The oxidation of sheep hemoglobin, in both the oxygenated and deoxygenated forms, by cuprous ions have been studied by spectrophotometric and stopped-flow techniques. Mixing of both the oxy and deoxy forms with excess Cu2+ leads to the rapid oxidation of the iron atoms of all four of the hem groups of the tetrameric protein, followed by the slow formation of hemichromes (low spin FeIII forms of hemoglobin). Stopped-flow studies show that the oxidations follow simple monophasic kinetics with second-order rate constants of 65 and 310 M-1 sec-1 for the oxy and deoxy forms, respectively. Variable temperature studies yield Arrhenius activation energies of 43 for the oxy form and 113 kJ mole-1 for the deoxy form. For each form of the protein the activation energy is very similar to the activation enthalpy. While the deoxy form is characterized by an activation energy and enthalpy that is more than twice the corresponding value in the oxy form. The activation entropies show highly significant differences being -128 e.u. and 136 e.u. at 25 degrees C for the oxy and deoxy forms, respectively.