Mechanistic Basis for Site–Site Interactions in Inhibitor and Substrate Binding to Band 3 (AE1): Evidence Distinguishing Allosteric from Electrostatic Effects

Salhany, James M.

doi:10.1006/bcmd.2001.0464

Cited by 13 publications

(13 citation statements)

References 36 publications

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“…For this purpose, DIDS, a selective inhibitor of anion transport that reacts covalently with Lys 539 of band 3 (46), was conjugated to biotin and employed to label a single band 3 molecule in intact cells. The specificity of the DIDS-biotin conjugate for mouse band 3 was established by reacting the conjugate with intact mouse erythrocytes, separating the component proteins by SDS-PAGE, blotting the proteins onto nitrocellulose paper, and visualizing the location of biotinylated polypeptides by staining with streptavidin-horseradish peroxidase.…”

Section: Resultsmentioning

confidence: 99%

Analysis of the Mobilities of Band 3 Populations Associated with Ankyrin Protein and Junctional Complexes in Intact Murine Erythrocytes

Kodippili

Spector

Hale

et al. 2012

Journal of Biological Chemistry

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Background: Erythrocyte band 3 exists in three populations; ankyrin-bound, adducin-bound, and free. Results: In wild-type murine erythrocytes, ϳ40% of band 3 is attached to ankyrin, ϳ33% is immobilized by adducin, and ϳ27% is free. Conclusion: Ankyrin-and adducin-bound band 3 can be monitored separately. Significance: This diffusion study demonstrates molecular differences between band 3 complexes and reveals structural heterogeneity within band 3 subpopulations.

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Section: Resultsmentioning

confidence: 99%

Analysis of the Mobilities of Band 3 Populations Associated with Ankyrin Protein and Junctional Complexes in Intact Murine Erythrocytes

Kodippili

Spector

Hale

et al. 2012

Journal of Biological Chemistry

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“…DIDS is a well-known AE1 inhibitor and reversibly binds to lysine 539 and lysine 851 of the exchanger (11). Thus, it is conceivable that lysine in albumin and in AE1 compete in binding DIDS, as albumin is rich in lysine.…”

Section: Effect Of Dids On No3mentioning

confidence: 99%

Effects of 4,4'-Diisothiocyanato-stilbene-2,2'-disulfonic Acid (DIDS) and Chlorpromazine on NO3− Transport via Anion Exchanger in Erythrocytes: Inertness of DIDS in Whole Blood

et al. 2003

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Abstract. We examined the effects of chlorpromazine on NO 3 -transport between erythrocytes (RBCs) and extracellular fluid. Chlorpromazine (10 m g / ml) did not influence NO 3 -movement in both whole blood and RBC suspension. Though an anion exchanger (AE1) inhibitor DIDS (4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid, 100 mM) did not alter NO 3 -movement in whole blood, it inhibited the movement in a concentration-dependent manner in the RBC suspension. The inhibition was abrogated by plasma and albumin concentration-dependently. Our results indicated that chlorpromazine had no effect on NO 3 -transport through AE1 and that the inertness of DIDS on AE1 in whole blood is due to interference by albumin in plasma. ) concentration has been used as an index of NO formation in vivo in spite of the high susceptibility to many confounding factors (1, 2). Among these factors, the possible accumulation of NO 3 -into erythrocytes (RBCs) through anion exchanger (AE1) has received attention as a modulating mechanism of plasma NO 3 -(almost all NOx in plasma) concentration (3), although the operation of this mechanism seems to be unlikely in healthy subjects (4). Recently, Herken et al. (5) reported higher NOx concentrations in washed RBCs of patients with schizophrenia, compared with control subjects. This result was inconsistent with our experience that the washing procedure could partially or completely reduce RBC NOx to a level equivalent to that in the washing solution. Therefore, it is difficult to accept the argument advanced by Herken et al. (5) that higher NOx concentration in the blood (hence in RBCs) reflects increment of NO formation in schizophrenia. However, a possible interpretation remains for the discrepancy. Since their patients were treated with neuroleptics (5), including chlorpromazine and haloperidol (6), it is possible that these agents may prevent NO 3 -transport through AE1. To test this hypothesis, we examined whether these agents could modify NO 3 -transport across the RBC cell membrane via AE1. In a previous study (4), we could not confirm the contribution of AE1 to NO 3 -transport in whole blood using 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid (DIDS), an AE1 inhibitor. Accordingly, experiments were performed using both whole blood and RBC suspension. We also examined the cause of inertness of DIDS in whole blood.Venous blood samples were drawn from healthy volunteers of both sexes (28 -50-year-old) after obtaining written consent. The study was approved by the Human Ethics Committee of Kanazawa Medical University. The blood sample was immediately heparinized and used directly or as RBC suspension. Phosphatebuffered saline (PBS; 10 mM, pH 7.4, NOx <0.1 m M) was prepared and RBCs were washed with five volumes of the buffer followed by centrifugation. The buffy coat

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“…The degree of echinocytosis by DNDS correlates with the degree of band 3 saturation without observing a spheroechinocytosis after band 3 saturation, indicating that the echinocytosis is due to the inhibition of the band 3 anion exchange. The inference of a stomatocytosis followed by an echinocytosis by a stilbenedisulfonic acid derivative is supported by previous stopped-flow kinetics of binding of covalently modifying DIDS or non-covalently modifying 4,4'-dibenzaminostil-bene-2,2'-disulfonic acid (DBDS) to band 3 in ghosts which show that there is initially a fast phase, followed by a slower phase of its binding, which are dependent and independent of their concentrations, respectively [22,23]. The inhibition of the band 3 transport of Cl -by a stilbenedisulfonic acid derivative is competitive although this derivative does not fully inhibit Cl -binding to the transport site, as indicated by Cl -binding to band 3 in ghosts by chloride-35 nuclear magnetic resonance (Cl 35 -NMR) spectroscopy [23,24].…”

mentioning

confidence: 79%

“…The inference of a stomatocytosis followed by an echinocytosis by a stilbenedisulfonic acid derivative is supported by previous stopped-flow kinetics of binding of covalently modifying DIDS or non-covalently modifying 4,4'-dibenzaminostil-bene-2,2'-disulfonic acid (DBDS) to band 3 in ghosts which show that there is initially a fast phase, followed by a slower phase of its binding, which are dependent and independent of their concentrations, respectively [22,23]. The inhibition of the band 3 transport of Cl -by a stilbenedisulfonic acid derivative is competitive although this derivative does not fully inhibit Cl -binding to the transport site, as indicated by Cl -binding to band 3 in ghosts by chloride-35 nuclear magnetic resonance (Cl 35 -NMR) spectroscopy [23,24]. It is attributed either to the binding of the stilbenedisulfonic acid derivative to the transport site [24] or to an allosteric site [23].…”

mentioning

confidence: 79%

“…The inhibition of the band 3 transport of Cl -by a stilbenedisulfonic acid derivative is competitive although this derivative does not fully inhibit Cl -binding to the transport site, as indicated by Cl -binding to band 3 in ghosts by chloride-35 nuclear magnetic resonance (Cl 35 -NMR) spectroscopy [23,24]. It is attributed either to the binding of the stilbenedisulfonic acid derivative to the transport site [24] or to an allosteric site [23]. However, the following observations would support the view that the derivative binds to the transport site.…”

mentioning

confidence: 99%

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A Basis of the Stomatocytoses of Erythrocytes by 1-Chloro-2, 4-Dinitrobenzene and Other Electrophilic Reagents

Wong¹

2015

Open Biol Sci J

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On bases of previous observations on the band 3 anion exchange inhibition by the electrophilic reagent 1-fluoro-2,4-dinitrobenzene in resealed ghosts and a previously proposed band 3-based mechanism of control of the erythrocyte shape, it is suggested that stomatocytoses by 1-chloro-2,4-dinitrobenzene and other electrophilic reagents are due to an inhibition of the band 3 anion exchange by a covalent modification of Lys 539, one of the two lysine intramolecularly cross-linked by impermeant 4,4'-diisothiocyanodihydrostilbene-2,2'-disulfonic acid, a band 3 anion exchange potent inhibitor. Attempts to explain these stomatocytoses led to explain the echinocytosis and competitive inhibition of band 3 anion exchange by stilbenedisulfonic acid derivatives and the reversal of the echinocytosis by 2,4-dinitrophenol by one stomatocytogenic of the electrophilic reagents. The inference on stomatocytoses by electrophilic reagents is of interest since stomatocytogenic and echinocytogenic by amphiphiles are perceived to interact noncovalently with the membrane and that they can be clinically relevant.

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Mechanistic Basis for Site–Site Interactions in Inhibitor and Substrate Binding to Band 3 (AE1): Evidence Distinguishing Allosteric from Electrostatic Effects

Cited by 13 publications

References 36 publications

Analysis of the Mobilities of Band 3 Populations Associated with Ankyrin Protein and Junctional Complexes in Intact Murine Erythrocytes

Analysis of the Mobilities of Band 3 Populations Associated with Ankyrin Protein and Junctional Complexes in Intact Murine Erythrocytes

Effects of 4,4'-Diisothiocyanato-stilbene-2,2'-disulfonic Acid (DIDS) and Chlorpromazine on NO3− Transport via Anion Exchanger in Erythrocytes: Inertness of DIDS in Whole Blood

A Basis of the Stomatocytoses of Erythrocytes by 1-Chloro-2, 4-Dinitrobenzene and Other Electrophilic Reagents

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