Haptoglobin--haemoglobin interaction. Heterogeneity of the haptoglobin 1-1 molecule in its binding affinity for horse R/E-hemoglobin.

The formation of two different complexes when haptoglobin (Hp) and haemoglobin (Hb) are mixed in a 1:1 molar ratio is demonstrated by isoelectrofocusing. In these two complexes, the affinity of Hp for Hb is shown to be different, since Hb can be displaced only from one of the complexes, by a further addition of Hp. This is confirmed by a quantitative study of the reaction stoichiometry, when [Hp]/[Hb] = 1 and [Hp]/[Hb] greater than 1, which allows an evaluation of the amount of each complex formed. All these data cannot be explained other than by the existence of two forms of Hp molecule and a reaction scheme which fits these experiments is proposed.