Light-scattering measurements of hemoglobin binding to the erythrocyte membrane. Evidence for transmembrane effects related to a disulfonic stilbene binding to band 3

Salhany, James M.; Cordes, Karen A.; Gaines, Elizabeth D.

doi:10.1021/bi00548a028

Cited by 141 publications

(69 citation statements)

References 34 publications

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“…Comparison of the direct binding affinity of deoxyHb for wild-type and 2 mutants of cdb3. Purified wild-type cdb3 (residues 1-379), a low-affinity mutant [del (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)] and a high-affinity mutant [del(29-52)] were immobilized onto Affi-Gel 15 beads. Derivatized beads were equilibrated in 10 mM bis-Tris acetate buffer, pH 6.5, and excess predialyzed Hb was added before incubation for 1 hour at room temperature with gentle agitation under argon.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, the anion transport inhibitor 4,4Ј-diisothiocyanatostilbene-2,2Ј-disulfonic acid (DIDS) is known to react almost exclusively with band 3 on (Lys539 or Lys542) near the outer surface of the erythrocyte membrane. 47,48 It is noteworthy that DIDS binding to this external site significantly alters Hb binding to the NH 2 terminus of cdb3, 18 suggesting that significant structural communication occurs between the deoxyHb binding site and the membranespanning domain of band 3. Could this conformational communication be used to signal the oxygenation state to solute transporters associated with the membrane-spanning domain of band 3?…”

Section: Discussionmentioning

confidence: 99%

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Characterization of the deoxyhemoglobin binding site on human erythrocyte band 3: implications for O2 regulation of erythrocyte properties

Chu

Breite

Ciraolo

et al. 2008

Blood

108

131

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Band 3, the major protein of the human erythrocyte membrane, associates with multiple metabolic, ion transport, and structural proteins. Functional studies demonstrate that the oxygenation state of the erythrocyte regulates cellular properties performed by these and/or related proteins. Because deoxyhemoglobin, but not oxyhemoglobin, binds band 3 reversibly with high affinity, these observations raise the hypothesis that hemoglobin might regulate erythrocyte properties through its reversible, oxygenationdependent association with band 3. To explore this hypothesis, we have characterized the binding site of deoxyHb on human erythrocyte band 3. We report that (1) deoxyHb binds to residues 12-23 of band 3; (2) mutation of residues on either side of this sequence greatly enhances affinity of deoxyHb for band 3, suggesting that evolution of a higher affinity interaction would have been possible had it been beneficial for survival; (3) Hb does not bind to 2 other sequences in band 3 despite their high sequence homology to residues 12-23, and (4) IntroductionSeveral lines of evidence suggest that the oxygenation state of the erythrocyte (RBC) regulates multiple erythrocyte properties. First, the activities of many membrane solute transporters change with the oxygen content of the cell, including volume regulatory transporters (eg, Na ϩ /H ϩ exchanger), cation coupled Cl Ϫ cotransporters, amino acid transporters, and ion channels. [1][2][3][4][5][6][7][8][9][10] The K/Cl cotransporter, for example, is reported to be 20-fold more active in oxygenated than in deoxygenated RBCs. 9,10 Second, erythrocyte metabolism is modulated by the O 2 tension of the medium. Thus, glucose flux through the pentose phosphate pathway proceeds twice as rapidly in oxygenated as in deoxygenated cells, and glucose consumption in glycolysis is inhibited by oxygenation. 11 Third, erythrocyte deoxygenation may affect membrane structural properties. For example, deoxygenated hemoglobin (Hb) (but not oxygenated Hb [oxyHb]) competes with ankyrin for binding to band 3, and band 3 retention in detergent-extracted membrane skeletons is greatly reduced in skeletal pellets from deoxygenated cells (M. Stefanovic and P.S.L., unpublished data, December 2005). Furthermore, ankyrin epitopes are more accessible to antiankyrin antibodies in deoxygenated than in oxygenated RBCs. Taken together, these latter findings imply a looser association of ankyrin with the membrane in deoxygenated than in oxygenated erythrocytes.In searching for a mechanism to account for oxygenation effects on red cell properties, identification of both the O 2 sensor and downstream effector was considered important. A major candidate for the O 2 sensor was presumed to be Hb, because the O 2 dependence of the above properties generally follows the O 2 dissociation curve of Hb. 4,12,13 The downstream effector has been more uncertain, but band 3 has represented the prime candidate, because it constitutes the only established binding site of Hb on the membrane. Moreover, multiple observation...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Characterization of the deoxyhemoglobin binding site on human erythrocyte band 3: implications for O2 regulation of erythrocyte properties

Chu

Breite

Ciraolo

et al. 2008

Blood

108

131

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“…At these concentrations, hemoglobin interacts with membrane proteins, binds to the cytoplasmic domain ofprotein 3 (1)(2)(3)(4)(5)(6)(7)(8), and stabilizes the self-association of spectrin, the major structural protein of the membrane skeleton (9). The modification of the erythrocyte membrane by abnormal hemoglobins is emerging as an important theme in the attempt to understand the pathophysiology of membrane damage in hemoglobinopathies and the process of erythrocyte senescence (10)(11)(12)(13)(14)(15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

Membrane protein lesions in erythrocytes with Heinz bodies.

Platt¹,

Falcone²

1988

J. Clin. Invest.

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We studied Heinz body-containing erythrocytes with three different unstable hemoglobins: Nottingham, Brockton, and unclassified. We demonstrated two classes of membrane protein defects in unstable hemoglobin-containing cells (UHRBCs), a defect of the spectrin-depleted inside-out vesicle (UH-IOV), and a defect of spectrin (UH-spectrin) itself.The composition of UH-IOVs is the same as control with respect to quantity of ankyrin and proportion inside-out. However, UH-IOVs bind even less spectrin than IOVs derived from sickle erythrocytes (SS-IOVs), suggesting a severe functional defect in the ankyrin of UH-RBCs (UH-ankyrin). Further evidence that UH-ankyrin is abnormal is demonstrated by the virtual absence of ankyrin in isotonic membrane shells of UH-RBCs (UH-shells), and abnormal mobility and decreased binding of the 72-kD (spectrin-binding) a-chymotryptic fragment of UH-ankyrin (UH-72-kD) to control spectrin.All UH-RBC membranes were spectrin-deficient (60% of control). In addition, spectrin isolated from UH-RBCs (UHspectrin) was abnormal in two respects: (a) presence of a fastmoving band on nondenaturing polyacrylamide gels of both 0°C and 37°C extracts, and (b) decreased binding to actin in the presence of protein 4

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“…The cytoplasmic pole of the membrane-spanning protein Band 3 has been identified as the high-Mfinity binding site for haemoglobin [1][2][3][4]. More recently, interaction of denatured haemoglobin derivatives with the erythrocyte membrane was also investigated.…”

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confidence: 99%

The distribution and aggregatability of intramembrane particles in phenylhydrazine-treated human erythrocytes

Lelkes

Fodor

Hollán

et al. 1988

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Freeze-fracture analysis of phenylhydrazine-treated, unfixed human erythroeytes showed a ramlom distribution of intramembran¢ particles both over membrane-bound Heinz.bodies and in the intervening areas when examined after fast freezing in liquid pr~,pane. The same results was obtained when unfixed, glyeerinated red cells were frozen in liquid Freon. In contrast to previously published data (Low et al. (1985) Science 227, 531-533) these results indicate that binding of Heinz-bodies to the red cell membrane cannot cause morphologically detectable clustering of Band 3 in phenylhydrazine-treated red cells. Over numerous Heinz-bodles a decreased Acridine orange-induced particle aggregation was observed. The phenomenon of the oxidant-induced red cell fluorescence is described.In recent years haemoglobin-erythrocyte membrane interaction has been the subject of considerable investigation. The cytoplasmic pole of the membrane-spanning protein Band 3 has been identified as the high-Mfinity binding site for haemoglobin [1][2][3][4]. More recently, interaction of denatured haemoglobin derivatives with the erythrocyte membrane was also investigated. Waugh and Low [5] reported that haemichromes induced by phenylhydrazine-treatment of haemoglobin have a much higher affinity for the cytoplasmic domain of Band 3 than native haemoglobin and that these haemichromes caused the isolated cytoplasmic segment of Band 3 to precipitate in solution.

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Light-scattering measurements of hemoglobin binding to the erythrocyte membrane. Evidence for transmembrane effects related to a disulfonic stilbene binding to band 3

Cited by 141 publications

References 34 publications

Characterization of the deoxyhemoglobin binding site on human erythrocyte band 3: implications for O2 regulation of erythrocyte properties

Characterization of the deoxyhemoglobin binding site on human erythrocyte band 3: implications for O2 regulation of erythrocyte properties

Membrane protein lesions in erythrocytes with Heinz bodies.

The distribution and aggregatability of intramembrane particles in phenylhydrazine-treated human erythrocytes

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