Sickle hemoglobin (HbS) is a point mutation of the two β subunits in normal Hb (HbA) that leads to nucleated polymerization and accompanying pathology. We measured the rates of homogeneous and heterogeneous nucleation of HbS in the presence of up to 50% HbA under conditions in which hybrid HbAS molecules will also form. The replacement of 50% of HbS by HbA slows polymerization by factors of ∼100 in the physiological range, which is substantially less than previously thought. To provide a theoretical description of these data, we extended the double nucleation model for HbS polymerization to conditions in which hybridized mixtures are present. Measurements of homogeneous nucleation and the theory agree only when at least one of the molecules in the nucleus is not a hybrid. We attribute this to the necessary presence in the nucleus of a molecule that utilizes both β-subunit mutation sites in intermolecular contacts, whereas the remaining molecules engage only one of the mutation sites. Heterogeneous nucleation appears to require an even greater number of nonhybrid molecules, presumably because of the need for the nucleus to attach to the polymer as well as to form internal bonds. These results also provide insights into the pathophysiology of sickle cell disease, including the occasional severe events that strike persons in whom both HbS and HbA are expressed, a condition known as sickle trait. The studies reported here are necessary for understanding physiologically relevant polymerization in the presence of ligands as well as therapeutically relevant copolymerizing inhibitors.
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