Partial characterization of the copolymerization reaction of erythrocyte membrane band 3 with hemichromes.
Early intermediates in the denaturation of hemoglobin, termed hemichromes, have been found previously to associate with the cytoplasmic domain of erythrocyte membrane band 3 in a manner which rapidly propagates into an insoluble, macroscopic copolymer. Because this interaction is thought to force a redistribution of band 3 in situ, the properties of the copolymerization reaction were investigated in greater detail. The band 3-hemichrome coaggregate was found to be stabilized largely by ionic interactions since elevation of either ionic strength or pH led to dissolution of the complex. The pH dependence, however, shifted to a more alkaline pH with increasing hemichrome concentration, suggesting a strong linkage between band 3 or hemichrome protonation and copolymer formation. The stoichiometry of the copolymer was measured at five globin chains per band 3 chain whenever underivatized dimer-tetramer hemichrome mixtures were employed. However, cross-linking of the hemichromes at either the alpha or the beta chains to form the stabilized tetramer yielded a copolymer stoichiometry of approximately eight globin chains per band 3 chain, i.e., two hemichrome sites per band 3 subunit. While underivatized hemichromes exhibited both a fast and slow phase of copolymerization, the cross-link-stabilized tetrameric hemichromes displayed predominantly the fast phase kinetics. Naturally occurring disulfide cross-linked hemichromes also reacted more avidly with band 3 than their reduced counterparts; however, the copolymerization process also proceeded to completion with totally reduced components. It is concluded that copolymerization of band 3 with hemichromes should occur under normal cellular conditions and at an accelerated velocity when the intracellular reducing power is low.
D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D004910:Erythrocyte Membrane; D006420:Hemeproteins / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046911:Macromolecular Substances; D009994:Osmolar Concentration
D001457:Anion Exchange Protein 1, Erythrocyte; D006420:Hemeproteins; D046911:Macromolecular Substances; C001332:hemichrome