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

Jacob, Harry S.; Brain, M. C.; Dacie, J. V.

doi:10.1172/jci105950

Cited by 76 publications

(28 citation statements)

References 26 publications

(32 reference statements)

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“…The mechanism by which glycolysis reduces membrane sulfhydryl groups (or possibly, mixed disulfides between hemoglobin and membrane sulfhydryl groups [14]), is not established by these experiments. It may be that this is achieved by reduced glutathione which can maintain hemoglobin and other proteins in the reduced state and depends itself upon glycolysis for reduction from the oxidized state.…”

Section: Discussionmentioning

confidence: 80%

Sulfhydryl Groups of the Erythrocyte Membrane and their Relation to Glycolysis and Drug-Induced Hemolytic Anemia

Zipursky¹,

Stephens²,

Brown³

et al. 1974

J. Clin. Invest.

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A B S T R A C T Hemolytic anemia caused by oxidative drugs is thought to result from the oxidation of-intracellular and membrane sulfhydryl groups of the erythrocyte. This process is more likely to occur in those erythrocytes in which the intracellular mechanism for reduction of disulfides is abnormal (e.g., glucose-6-phosphate dehydrogenase deficiency). If a membrane sulfhydryl group is critical inthe pathogenesis of druginduced henioly;tic anemia, it follows that this specific group must be dependent on intracellular reductive mechanisms for maintenance of the reduced state.This report describes a sulfhydryl group(s), involved in membrane structure, which is (iare) dependent on intracellular metabolism for maintenance of the reduced state. It is postulated that this metabolically dependent membrane sulfhydryl group may play a role in the pathogenesis of drug-induced hemolytic anemia.Membrane sulfhydryl groups were studied by observing the effect of sullfhydryl blocking agents, e.g., N-ethylmaleimide (NEM), on the recovery of erythrocyte ghosts after osmotic lysis. It was shown that NEM interfered with ghost recovery by reacting with membrane sulfhydryl groups. The concentration of NEM (as determined by [1"C] NEM binding) necessary to cause this effect was lower than that necessary to produce changes in osmotic fragility or cation permeability, or to cause Heinz body formation.In the absence of glucose, these sulfhydryl groups became disulfides, but could be returned to the reduced state by restoring glycolysis or by adding dithiothreitol.Phenyihydrazine hemolytic anemia was induced in pigs, and membrtane changes of the type described above occurred early in the pathogenesis of the disease.

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Section: Discussionmentioning

confidence: 80%

Sulfhydryl Groups of the Erythrocyte Membrane and their Relation to Glycolysis and Drug-Induced Hemolytic Anemia

Zipursky¹,

Stephens²,

Brown³

et al. 1974

J. Clin. Invest.

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“…However, in unstable Hb disease the components of the disulfide-bonded peptide material were different from that found in G6PD mutants in that globin was a principal constituent. (4), and more recent evidence that Hb is reversibly bound to band 3 (23), to which it can become disulfide-linked by oxidation (24).…”

Section: Discussionmentioning

confidence: 99%

Oxidant damage of the lipids and proteins of the erythrocyte membranes in unstable hemoglobin disease. Evidence for the role of lipid peroxidation.

Flynn¹,

Allen²,

Johnson³

et al. 1983

J. Clin. Invest.

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A B S T R A C T Since unstable hemoglobins have been considered a source of reactive oxygen radicals, and oxidative membrane damage a prehemolytic event,we examined the erythrocyte membranes of six patients (three splenectomized) with hemoglobin Koln disease. In the hydrogen peroxide stress test, the patients' erythrocytes generated more than twice the malonyldialdehyde (a lipid peroxidative product) than control erythrocytes. Fluorescence spectra of lipid extracts of the patients' erythrocytes showed an excitation maximum at 400 nm and an emission maximum of 460 nm, characteristic of malonyldialdehyde lipid adducts. Two types of membrane polypeptide aggregates were found in the erythrocytes of the splenectomized patients. The first, which were dissociable by treatment with mercaptoethanol, contained disulfidelinked spectrin, band 3 and globin. The second, not dissociable by mercaptoethanol, had an amino acid composition similar to that of erythrocyte membranes and spectrin (unlike globin) and like that of aggregates produced by the action of malonyldialdehyde on normal erythrocyte membranes. Atomic absorption spectroscopy of hemoglobin Koln erythrocytes showed no increase in calcium content implying that these cross-links were not due to calcium-stimulated trans aggregates from splenectomized patients were less deformable while aggregate-free erythrocytes from nonsplenectomized patients had normal deformability. We conclude that the erythrocyte membranes in hemoglobin Koln disease show evidence of lipid peroxidation with production of malonyldialdehyde, and that the nondissociable membrane aggregates formed in this disease are likely cross-linked by malonyldialdehyde. Because the erythrocytes containing membrane aggregates from splenectomized patients with unstable hemoglobin disease show decreased membrane deformability, we hypothesize that this abnormality results in premature erythrocyte destruction in vivo.

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“…Isolated CHBHA hemoglobins and a2lemepao were incubated as 0.01 M phosphate-buffered (pH 7.4) solutions either at 37°C or 50°C. The latter temperature leads to precipitation of the mutant unstable hemoglobins into typical coccoid Heinz bodies (9), which underlies its use in the convenient screening procedure for CHBHA (1). Following centrifugation at 25,000 g, heme losses from the supernatant solutions of incubated hemo-1 Exceptions include hemoglobins Torinoa43 phenylalanine-valine in which a mutation occurs in close apposition to the a-chain heme group at the homologous site as that found in the a-chain of hemoglobin Hammersmith (12), and hemoglobins RiverdaleBronx#24 glycine-.arginine (13), PhillyP35 tyrosine-.phenylalanine (14), and Bibbaal36 leucine-oproline (15).…”

Section: Methodsmentioning

confidence: 99%

“…Precipitated hemoglobin was quantitated by assaying the difference in 280 mA absorbency before and after incubation; supernatant solutions from centrifuged (25,000 g) samples were utilized. Alternatively gravimetric analysis was utilized as previously described (9). For the latter the precipitate from a known quantity of hemoglobin was collected by centrifugation at 25,000 g, washed twice with distilled water, and dried to constant weight in vacuo over P205; the method was similar to that used by Jandl, Engle, and Allen (26).…”

Section: Methodsmentioning

confidence: 99%

“…From evidence that hemoglobin Koln binds its heme groups less avidly than hemoglobin A, one of us has suggested that heme groups might inordinately be lost from mutant globin polypeptide chains of CHBHA hemoglobins, predisposing them to denaturation into Heinz bodies (8,9). It was emphasized that the amino acid alteration in K6ln, as well as three other unstable hemoglobins under study at that time, Hammersmith (6), Zurich (7), and Gun Hill (10), were all in the vicinity of the heme group of the beta chain of hemoglobin.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The role of hemoglobin heme loss in Heinz body formation: studies with a partially heme-deficient hemoglobin and with genetically unstable hemoglobins

Jacob

Winterhalter²

1970

J. Clin. Invest.

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

Cited by 76 publications

References 26 publications

Sulfhydryl Groups of the Erythrocyte Membrane and their Relation to Glycolysis and Drug-Induced Hemolytic Anemia

Sulfhydryl Groups of the Erythrocyte Membrane and their Relation to Glycolysis and Drug-Induced Hemolytic Anemia

Oxidant damage of the lipids and proteins of the erythrocyte membranes in unstable hemoglobin disease. Evidence for the role of lipid peroxidation.

The role of hemoglobin heme loss in Heinz body formation: studies with a partially heme-deficient hemoglobin and with genetically unstable hemoglobins

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