Effect of organic co-solvents on the dimer/tetramer equilibrium of human haemoglobin.

We have studied the effect of organic co-solvents (monohydric alcohols and formamide) on the dimer-tetramer equilibrium of human haemoglobin by measuring the dependence of oxygen affinity upon haemoglobin concentration over a 100-fold concentration range and analysing the data with a modified Monod-Wyman-Changeux model in which the equilibrium (tetramer) in equilibrium with (dimer) + (dimer) was taken into account. This procedure enabled us to obtain the dimer-tetramer equilibrium constant and to find its dependence upon co-solvent concentration. Then by following the procedure already reported for the tense----relaxed state transition of haemoglobin, we separated the co-solvent effects into bulk electrostatic and non-bulk electrostatic (hydrophobic) contributions. We believe that our results demonstrate that during haemoglobin dissociation, just as we have already shown for the tense to relaxed state transition, both charged groups and hydrophobic surfaces became exposed to the solvent.