Further Studies on Zn2+-Mediated Domain–Domain Communication in Human Erythrocyte Band 3

1998 ◽  
Vol 18 (5) ◽  
pp. 265-277
Author(s):  
Hong Xu ◽  
Xujia Zhang ◽  
Fu Yu Yang
Keyword(s):  
Conformational Change ◽  
Human Erythrocyte ◽  
Cytoplasmic Domain ◽  
Band 3 ◽  
Anion Transport ◽  
Intrinsic Fluorescence ◽  
Transport Activity ◽  
Membrane Domain ◽  
Nmr Study ◽  
Anion Transport Activity

Human erythrocyte band 3 is purified and reconstituted into vesicles, forming right-side-out proteoliposomes. Zn2+ entrapped inside the proteoliposomes inhibits the anion transport activity of band 3, and removal of the cytoplasmic domain of band 3 is able to diminish Zn2+ inhibition. Thus, the inhibition of activity of band 3 results from the Zn2+ induced conformational change of the cytoplasmic domain, which in turn is transmitted to the membrane domain. The results of intrinsic fluorescence and its quenching by HB and the 35Cl NMR study indicate that the cytoplasmic domain is essential for the conformational change induced by Zn2+.SH-blocking reagents, CH3I and GSSG, are used to modify the cytoplasmic domain, where they specifically bind to Cys201 and Cys317. It is observed that the Zn2+ induced inhibition of anion transport activity is blocked. This demonstrates that Cys201 and Cys317 are required in Zn2+-mediated domain–domain communication.

scholarly journals Band 3 HT, a human red-cell variant associated with acanthocytosis and increased anion transport, carries the mutation Pro-868→Leu in the membrane domain of band 3

1993 ◽  
Vol 293 (2) ◽  
pp. 317-320 ◽  
Author(s):  
L J Bruce ◽  
M M Kay ◽  
C Lawrence ◽  
M J Tanner
Keyword(s):  
Cell Shape ◽  
Band 3 ◽  
Anion Transport ◽  
Red Cell ◽  
Transport Activity ◽  
Binding Properties ◽  
Red Cell Shape ◽  
Disulphonic Acid ◽  
Anion Transport Activity ◽  
Cell Variant

1. We have studied band 3 HT, a human red-cell band 3 variant with increased M(r), which is associated with abnormal red-cell shape (acanthocytosis) and increased anion-transport activity. 2. We have shown that the increased M(r) does not result from the presence of the band 3 Memphis mutation, and that the variant band 3 is covalently labelled by 4,4′-di-isothiocyanato-1,2-diphenylethane-2,2′-disulphonic acid (H2DIDS) less readily than normal. 3. cDNA cloning studies show that band 3 HT results from the mutation Pro-868-->Leu, and the possible significance of the mutation in the altered anion-transport activity and cytoskeleton binding properties of band 3 HT is discussed.

Exhaustive-exercise-induced oxidative stress alteration of erythrocyte oxygen release capacity

2018 ◽  
Vol 96 (9) ◽  
pp. 953-962 ◽  
Author(s):  
Yanlian Xiong ◽  
Yanlei Xiong ◽  
Yueming Wang ◽  
Yajin Zhao ◽  
Yaojin Li ◽  
...  
Keyword(s):  
Carbonic Anhydrase ◽  
Band 3 ◽  
Anion Transport ◽  
Kinetic Properties ◽  
Oxygen Release ◽  
Transport Activity ◽  
Running Exercise ◽  
Release Capacity ◽  
Anion Transport Activity ◽  
2,3 Dpg

The aim of the present study was to explore the effect of exhaustive running exercise in the oxygen release capacity of rat erythrocytes. Rats were divided into sedentary control, moderate running exercise, and exhaustive running exercise groups. The thermodynamic and kinetic properties of the erythrocyte oxygen release process of the different groups were tested. We also determined the degree of band-3 oxidation and phosphorylation, anion transport activity, and carbonic anhydrase isoform II activity. Biochemical studies suggested that exhaustive running significantly increased oxidative injury parameters in thiobarbituric acid reactive substances and methaemoglobin levels. Furthermore, exhaustive running significantly decreased anion transport activity and carbonic anhydrase isoform II activity. Thermodynamic analysis indicated that erythrocytes oxygen release ability also significantly increased due to elevated 2,3-DPG level after exhaustive running. Kinetic analysis indicated that exhaustive running resulted in significantly decreased T50 value. We presented evidence that exhaustive running remarkably impacted thermodynamic and kinetic properties of RBC oxygen release. In addition, changes in 2,3-DPG levels and band-3 oxidation and phosphorylation could be the driving force for exhaustive-running-induced alterations in erythrocyte oxygen release thermodynamic and kinetic properties.

scholarly journals Transport activity of chimaeric AE2-AE3 chloride/bicarbonate anion exchange proteins

2003 ◽  
Vol 371 (3) ◽  
pp. 687-696 ◽  
Author(s):  
Jocelyne FUJINAGA ◽  
Frederick B. LOISELLE ◽  
Joseph R. CASEY
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Cytoplasmic Domain ◽  
Anion Transport ◽  
Transport Activity ◽  
Surface Processing ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
Anion Transport Activity

Chloride/bicarbonate anion exchangers (AEs), found in the plasma membrane of most mammalian cells, are involved in pH regulation and bicarbonate metabolism. Although AE2 and AE3 are highly similar in sequence, AE2-transport activity was 10-fold higher than AE3 (41 versus 4 mM · min−1 respectively), when expressed by transient transfection of HEK-293 cells. AE2–AE3 chimaeras were constructed to define the region responsible for differences in transport activity. The level of AE2 expression was approx. 30% higher than that of AE3. Processing to the cell surface, studied by chemical labelling and confocal microscopy, showed that AE2 is processed to the cell surface approx. 8-fold more efficiently than AE3. The efficiency of cell-surface processing was dependent on the cytoplasmic domain, since the AE2 domain conferred efficient processing upon the AE3 membrane domain, with a predominant role for amino acids 322–677 of AE2. AE2 that was expressed in HEK-293 cells was glycosylated, but little of AE3 was. However, AE2 expressed in the presence of the glycosylation inhibitor, tunicamycin, was not glycosylated, yet retained 85 ± 8% of anion-transport activity. Therefore glycosylation has little, if any, role in the cell-surface processing or activity of AE2 or AE3. We conclude that the low anion-transport activity of AE3 in HEK-293 cells is due to low level processing to the plasma membrane, possibly owing to protein interactions with the AE3 cytoplasmic domain.

scholarly journals Altered band 3 structure and function in glycophorin A- and B-deficient (MkMk) red blood cells

Blood ◽  
1994 ◽  
Vol 84 (3) ◽  
pp. 916-922 ◽  
Author(s):  
LJ Bruce ◽  
JD Groves ◽  
Y Okubo ◽  
B Thilaganathan ◽  
MJ Tanner
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Band 3 ◽  
Anion Transport ◽  
Glycophorin A ◽  
Sulfate Transport ◽  
Transport Activity ◽  
Anion Transporter ◽  
Glycan Chain ◽  
Anion Transport Activity

The anion transport activity of the human erythrocyte anion transporter (band 3; AE1) has been examined in both normal and glycophorin A (GPA)- deficient (MkMk) human red blood cells (RBCs). The sulfate transport activity of MkMk cells (from two ethnically diverse sources) was approximately 60% that of normal erythrocytes under the transport assay conditions used. However, MkMk and normal RBCs contained similar amounts of band 3. The reduction in sulfate transport activity was shown to be caused by an increase in the apparent Km for sulfate in MkMk RBCs, suggesting the band 3 in the MkMk RBCs has a lowered binding affinity for sulfate anions. The size of the N-glycan chain on band 3 of the MkMk cells was larger than that on band 3 from normal RBCs. In contrast, the size of the N-glycan chain on the glucose transporter (GLUT1) from MkMk cells was smaller than that on GLUT1 from normal cells. The possible role of GPA in the biosynthesis and anion transport activity of band 3 in normal RBCs is discussed.

Zn2+ inhibits the anion transport activity of Band 3 by binding to its cytoplasmic tail

1994 ◽  
Vol 14 (4) ◽  
pp. 159-169 ◽  
Author(s):  
Y. P. Tu ◽  
H. Xu
Keyword(s):  
Divalent Cations ◽  
Membrane Vesicles ◽  
Band 3 ◽  
Anion Transport ◽  
Dependent Manner ◽  
Transport Activity ◽  
Human Erythrocyte Membrane ◽  
Anion Transport Activity ◽  
Dose Dependent ◽  
Cytoplasmic Side

Zn2+ can induce a conformational change of Band 3 with concomitant inhibition of its anion transport activity of human erythrocyte membrane vesicles only from the cytoplasmic side. The Zn2+ inhibition exhibits a dose-dependent manner with an apparent half maximal concentration of 50 μM ZnCl2 and can be reversed by 0.5 mM EDTA, but not by 1 mM dithiothreitol. The Zn2+ effect is specific and no similar inhibitory action could be observed by other divalent cations (Cu2+, Mn2+, Mg2+ or Sr2+).

scholarly journals Altered band 3 structure and function in glycophorin A- and B-deficient (MkMk) red blood cells

Blood ◽  
1994 ◽  
Vol 84 (3) ◽  
pp. 916-922 ◽  
Author(s):  
LJ Bruce ◽  
JD Groves ◽  
Y Okubo ◽  
B Thilaganathan ◽  
MJ Tanner
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Band 3 ◽  
Anion Transport ◽  
Glycophorin A ◽  
Sulfate Transport ◽  
Transport Activity ◽  
Anion Transporter ◽  
Glycan Chain ◽  
Anion Transport Activity

Abstract The anion transport activity of the human erythrocyte anion transporter (band 3; AE1) has been examined in both normal and glycophorin A (GPA)- deficient (MkMk) human red blood cells (RBCs). The sulfate transport activity of MkMk cells (from two ethnically diverse sources) was approximately 60% that of normal erythrocytes under the transport assay conditions used. However, MkMk and normal RBCs contained similar amounts of band 3. The reduction in sulfate transport activity was shown to be caused by an increase in the apparent Km for sulfate in MkMk RBCs, suggesting the band 3 in the MkMk RBCs has a lowered binding affinity for sulfate anions. The size of the N-glycan chain on band 3 of the MkMk cells was larger than that on band 3 from normal RBCs. In contrast, the size of the N-glycan chain on the glucose transporter (GLUT1) from MkMk cells was smaller than that on GLUT1 from normal cells. The possible role of GPA in the biosynthesis and anion transport activity of band 3 in normal RBCs is discussed.

As2O3enhances the anion transport activity of band 3 and the action is related with the C-terminal 16 residues of the protein

2005 ◽  
Vol 13 (4) ◽  
pp. 235-243 ◽  
Author(s):  
Guo-Hui Fu ◽  
Yong Wang ◽  
Yu-Hui Xi ◽  
Zhuo-Wei Guo ◽  
Xiao-Bin Liu ◽  
...  
Keyword(s):  
Band 3 ◽  
Anion Transport ◽  
Transport Activity ◽  
Anion Transport Activity

Transmembrane Ca2+ gradient is essential for high anion transport activity of human erythrocytes

1996 ◽  
Vol 16 (4) ◽  
pp. 299-311 ◽  
Author(s):  
Y. P. Tu ◽  
C. Feng ◽  
H. Xu ◽  
Z. Y. Guang ◽  
Q. W. Lu ◽  
...  
Keyword(s):  
Band 3 ◽  
Anion Transport ◽  
Ionophore A23187 ◽  
Half Maximum ◽  
Erythrocyte Ghosts ◽  
Transport Activity ◽  
Maximum Inhibition ◽  
Anion Transport Activity ◽  
Resealed Ghosts

The role of a transmembrane Ca2+ gradient in anion transport by Band 3 of human resealed erythrocyte ghosts has been studied. The results show that a transmembrane Ca2+ gradient is essential for the conformation of erythrocyte Band 3 with higher anion transport activity. The dissipation of the transmembrane Ca2+ gradient by the ionophore A23187 inhibits the anion transport activity. The extent of this inhibition approaches 90% as the Ca2+ concentration on both sides of the ghost membrane is increased to 1.0 mM and half-maximum inhibition is observed at 0.25 mM Ca2+. Addition of ATP (0.4 mM) to the resealing medium can partly reestablish the transmembrane Ca2+ gradient by activation of Ca2+-ATPase and alleviate the inhibition to some extent. N-ethylmaleimide, an inhibitor of erythrocyte Ca2+-ATPase, prevents such restoration. Electron micrographs reveal that numerous larger intramembranous particles can be observed on the P-faces of freeze-fractured resealed ghosts in the absence of a transmembrane Ca2+ gradient.

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