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. 1968 Dec;47(12):2664–2677. doi: 10.1172/JCI105950

Altered sulfhydryl reactivity of hemoglobins and red blood cell membranes in congenital heinz body hemolytic anemia

Harry S Jacob 1,2, Michael C Brain 1,2, John V Dacie 1,2
PMCID: PMC297438  PMID: 5725279

Abstract

The mechanisms of hemoglobin precipitation into Heinz bodies and hemolytic anemia that characterize congenital Heinz body hemolytic anemia (CHBHA) were studied in patients with the unstable hemoglobins, Köln (β-98 valine → methionine) and Hammersmith (β-42 phenylalanine → serine). The cysteines in the 93rd position of the β-chains of CHBHA hemoglobins bound glutathione excessively in mixed disulfide linkage. The resulting diminished “free” GSH within the cell accelerated hexose monophosphate shunt metabolism. The unique precipitability of CHBHA hemoglobins when heated at 50°C could be induced in normal hemoglobin A by artificially blockading its sulfhydryl groups with paramercuribenzoate (PMB).

Reflecting the previously reported excessive flux of hemes from hemoglobin Köln, the expected heme/globin ratio in this hemoglobin was reduced by 30%. The further increment in heme loss that occurs with heat (50°C) underlies the unique heat precipitability of CHBHA hemoglobins; it was retarded if detachment of heme was inhibited by cyanide or carbon monoxide.

Heinz bodies were attached to red cell membrane thiol groups presumably through mixed disulfide bonds, being released by mercaptoethanol. Binding of hemoglobin Köln-59Fe to red cell ghosts, which was markedly enhanced when Heinz bodies were generated at 50°C, was inhibited if membrane thiols were preblockaded by PMB. The depletion of membrane thiols by their reaction with Heinz bodies rendered CHBHA red cells hypersusceptible to membrane sulfhydryl inhibitors, as manifested by inordinate cation leakage, osmotic fragility, and autohemolysis.

We conclude that both cellular and membrane thiols bind β-93 sulfhydryls of CHBHA hemoglobins as mixed disulfides. Concomitantly, heme avidity to β-92 lessens, suggesting that degradation of the resulting excessively freed heme may produce the pigmented dipyrroluria of this syndrome. Heinz bodies, reflecting the heightend precipitability of heme-deficient globin, attach to, thereby depleting, membrane sulfhydryl groups. This, as shown previously, could underlie the hemolytic anemia of this syndrome by causing membrane hyperpermeability, premature splenic entrapment, and ultimately osmotic destruction of red blood cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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