Interactions between band 3 and other transport-related proteins

A Cl(-)-HCO3- exchanger in the brush-border membrane mediates active Cl- absorption and regulates intracellular pH in rabbit ileum. The molecular identity of the ileal Cl(-)-HCO3- exchanger has not been established. The best-characterized plasma membrane Cl(-)-HCO3- exchanger is erythroid band 3. Structurally related proteins in nonerythroid tissues comprise an anion exchanger (AE) family. We used the polymerase chain reaction to amplify and clone a cDNA encoding an ileal band 3-related protein (B3RP) from rabbit ileal enterocytes. The composite sequence is 3,909 bp and is predicted to encode a protein of 136 kDa. The deduced amino acid sequence is 95% identical to murine renal AE2, indicating that ileal B3RP is rabbit AE2. Antisera generated against a cytoplasmic fragment of ileal B3RP recognized a 160- to 170-kDa polypeptide in the brush-border membrane, but not the basolateral membrane, of ileal crypt and villus enterocytes. This correlates with previous studies indicating that a Cl(-)-HCO3- exchange is present in brush-border but not basolateral membrane vesicles from rabbit ileal enterocytes. We conclude that ileal B3RP is a product of the AE gene family, and is present in the brush-border of ileal enterocytes, where it may mediate Cl(-)-HCO3- exchange.

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

Blood ◽

10.1182/blood-2007-07-100180 ◽

2008 ◽

Vol 111 (2) ◽

pp. 932-938 ◽

Cited By ~ 71

Author(s):

Haiyan Chu ◽

Andrew Breite ◽

Peter Ciraolo ◽

Robert S. Franco ◽

Philip S. Low

Keyword(s):

Binding Site ◽

Human Erythrocyte ◽

Band 3 ◽

Major Protein ◽

Human Erythrocyte Membrane ◽

Functional Studies ◽

High Sequence Homology ◽

Affinity Interaction ◽

Related Proteins

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, oxygenation-dependent 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) the Hb binding site on band 3 lies proximal to binding sites for glycolytic enzymes, band 4.1 and ankyrin, suggesting possible mechanisms through which multifarious erythrocyte properties might be regulated by the oxygenation state of the cell.

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Immunochemical characterization of a band 3-like anion exchanger in collecting duct of human kidney

AJP Renal Physiology ◽

10.1152/ajprenal.1987.253.2.f213 ◽

1987 ◽

Vol 253 (2) ◽

pp. F213-F221 ◽

Cited By ~ 10

Author(s):

S. Wagner ◽

R. Vogel ◽

R. Lietzke ◽

R. Koob ◽

D. Drenckhahn

Keyword(s):

Posttranslational Modifications ◽

Anion Exchanger ◽

Collecting Duct ◽

Basolateral Membrane ◽

Human Kidney ◽

Cytoplasmic Domain ◽

Band 3 ◽

Intercalated Cells ◽

Membrane Spanning ◽

Related Proteins

Poly- and monoclonal antibodies have been prepared against the cytoplasmic domain (43 kDa) and the 17-, 20-, and 35-kDa fragments of the membrane-spanning domain of the human erythrocyte anion exchanger, band 3. The antibodies were used to localize and further characterize analogues of band 3 in the human kidney. We report here that the basolateral membrane of intercalated cells of the connecting tubules and collecting ducts contains an analogue of band 3 that appears to be highly homologous to the erythrocyte anion exchanger. This band 3-like protein is probably important for reabsorption of bicarbonate in the collecting duct system and thus for acidification of the forming urine. The band 3-like protein of the intercalated cells contain immunoreactive sites of both the cytoplasmic domain and the three major fragments of the membrane-spanning domain of erythrocyte band 3. Although no immunological differences were detected between the membrane-spanning domains of band 3 in erythrocytes and intercalated cells, there are at least three sites along the cytoplasmic domain of kidney band 3 that differ from erythrocyte band 3 in either amino acid composition or posttranslational modifications. The main kidney analogue of band 3 that contains epitopes of the cytoplasmic domain as well as the 17- and 35-kDa membrane-spanning domain of erythroid band 3 is a polypeptide with an apparent molecular mass of 100-110 kDa. Further immunoreactive polypeptides at approximately 180, approximately 140, approximately 38, approximately 25-30 kDa that were detected at lower stringency and higher sensitivity of the immunoblotting procedure may be members of a multigene family that encodes a series of related proteins.

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Eosin-5-maleimide binding to band 3 and Rh-related proteins forms the basis of a screening test for hereditary spherocytosis

British Journal of Haematology ◽

10.1046/j.1365-2141.2003.04730.x ◽

2003 ◽

Vol 124 (1) ◽

pp. 106-113 ◽

Cited By ~ 58

Author(s):

May-Jean King ◽

Jonathan S. Smythe ◽

Rosey Mushens

Keyword(s):

Screening Test ◽

Hereditary Spherocytosis ◽

Band 3 ◽

Related Proteins

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Hereditary Spherocytosis: How the New Information Provided by the Routine Hematology Analysers May Help in Its Differential Diagnosis or Flagging

Blood ◽

10.1182/blood.v120.21.5163.5163 ◽

2012 ◽

Vol 120 (21) ◽

pp. 5163-5163

Author(s):

Susanna Barella ◽

Ramon Simon-Lopez ◽

Nicola Di Gaetano ◽

Renzo Galanello

Keyword(s):

Research Funding ◽

Roc Analysis ◽

Hereditary Spherocytosis ◽

Osmotic Fragility ◽

Peripheral Blood Smear ◽

Band 3 ◽

Screening Tests ◽

New Information ◽

Related Proteins ◽

Best Parameters

Abstract Abstract 5163 Introduction: Hereditary Spherocytosis (HS) is one of the most common inherited hemolytic anemias. Many of them are autosomal dominant, being about 25% of the cases transmitted recessively. Diagnostic of HS: Classic testing for HS includes: Hematologic testing of red blood cell indices (RDW, MCHC, Reticulocyte count), peripheral blood smear (presence of spherocytes), osmotic fragility and Eosin-5-maleimide binding to band 3 and Rh-related proteins forms that may be used as screening tests for hereditary spherocytosis. Objective: Recently have been developed new parameters and information in the new automated hematology analyzer called DxH8008™ from Beckman Coulter as @MSCV (@Mean Sphered Cell Volume), @RSF, @MAF, @ LHD%. All this parameters may be used to create flagging for laboratory use only (LUO) or Research use only (RUO). The purpose of this study is to investigate the possible use or utility of this new information for the screening/flagging of Hereditary Spherocytosis. There are previous studies showing the possible benefit of using MCV minus @MSCV for the detection/flagging of cases with spherocytes. Patient and Methods: We have collected 28 patients with Hereditary Spherocytosis. All of them were confirmed by red cell morphology, osmotic fragility and Eosin-5-maleimide binding to band 3 and Rh-related proteins forms. Results: Using ROC analysis, the best parameters differentiating the Hereditary Spherocytosis from the normals were: RET% (AUC 0. 996), MCV - @MSCV (AUC 0. 996), @MSCV (AUC 0. 969), RDW(AUC 0. 892), MCHC (AUC 0. 860), HGB (AUC 0. 787). Using ROC analysis, the best parameters differentiating the Hereditary Spherocytosis from other anemias (excluding normals)were: MCV - @MSCV (AUC 0. 991), MCHC (AUC 0. 987), RET% (AUC 0. 857). Disclosures: Simon-Lopez: Beckman Coulter: @LHD, @MAF, @RSF, @LHD, @MAF, @RSF Patents & Royalties, Employment. Di Gaetano:Instrumentation Laboratory spa: Work for a distributor of Beckman Coulter Instruments in Italy Other. Galanello:Novartis: Research Funding, Speakers Bureau; Apopharma: Research Funding, Speakers Bureau; Ferrokin: Research Funding.

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Anion Exchange Mechanism of Band 3 and Related Proteins

Membrane Transport in Biology ◽

10.1007/978-3-642-76983-2_5 ◽

1992 ◽

pp. 233-261 ◽

Cited By ~ 1

Author(s):

R. B. Gunn

Keyword(s):

Anion Exchange ◽

Band 3 ◽

Exchange Mechanism ◽

Related Proteins

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Band 3 Memphis: a widespread polymorphism with abnormal electrophoretic mobility of erythrocyte band 3 protein caused by substitution AAG---- GAG (Lys----Glu) in codon 56

Blood ◽

10.1182/blood.v80.6.1592.1592 ◽

1992 ◽

Vol 80 (6) ◽

pp. 1592-1598 ◽

Cited By ~ 41

Author(s):

P Jarolim ◽

HL Rubin ◽

S Zhai ◽

KE Sahr ◽

SC Liu ◽

...

Keyword(s):

Fusion Protein ◽

Glutamic Acid ◽

Electrophoretic Mobility ◽

Band 3 ◽

Normal Band ◽

Base Substitution ◽

Band 3 Protein ◽

The Difference ◽

Related Proteins ◽

Reduced Mobility

Abstract Band 3 Memphis (b3M) is a variant of the erythrocyte band 3 protein detected in individuals of virtually all ethnic groups and characterized by a reduced mobility of proteolytic fragments derived from the N-terminus of the cytoplasmic domain of band 3 (cdb3). We have sequenced band 3 cDNA corresponding to cdb3 in 12 heterozygotes for the b3M polymorphism including one white, one black, one Chinese, one Philippino, one Malay, and seven Melanesian subjects. In all individuals, we found a single-base substitution in codon 56 of one band 3 allele changing lysine to glutamic acid (AAG----GAG) which, in some of them, was linked with an additional mutation in cdb3. Since the change of codon 56 from AAG to GAG was the only mutation in the studied individuals found within the cDNA segment coding for the abnormally migrating fragment of cdb3, we conclude that it represents the underlying molecular basis of the b3M polymorphism. We further support this conclusion by showing that electrophoresis in the presence of 4 mol/L urea abolished the difference in migration between proteolytic products of b3M and normal band 3, and that a fusion protein prepared from cDNA coding for the b3M allele again exhibits reduced electrophoretic mobility compared with the normal fusion protein. Finally, since most of the previously cloned mouse, rat, and chicken band 3 and band 3-related proteins contain glutamic acid in the position corresponding to amino acid 56 in the human band 3, we propose that the Memphis variant is the evolutionarily older form of band 3.

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Band 3 Memphis: a widespread polymorphism with abnormal electrophoretic mobility of erythrocyte band 3 protein caused by substitution AAG---- GAG (Lys----Glu) in codon 56

Blood ◽

10.1182/blood.v80.6.1592.bloodjournal8061592 ◽

1992 ◽

Vol 80 (6) ◽

pp. 1592-1598

Author(s):

P Jarolim ◽

HL Rubin ◽

S Zhai ◽

KE Sahr ◽

SC Liu ◽

...

Keyword(s):

Fusion Protein ◽

Glutamic Acid ◽

Electrophoretic Mobility ◽

Band 3 ◽

Normal Band ◽

Base Substitution ◽

Band 3 Protein ◽

The Difference ◽

Related Proteins ◽

Reduced Mobility

Band 3 Memphis (b3M) is a variant of the erythrocyte band 3 protein detected in individuals of virtually all ethnic groups and characterized by a reduced mobility of proteolytic fragments derived from the N-terminus of the cytoplasmic domain of band 3 (cdb3). We have sequenced band 3 cDNA corresponding to cdb3 in 12 heterozygotes for the b3M polymorphism including one white, one black, one Chinese, one Philippino, one Malay, and seven Melanesian subjects. In all individuals, we found a single-base substitution in codon 56 of one band 3 allele changing lysine to glutamic acid (AAG----GAG) which, in some of them, was linked with an additional mutation in cdb3. Since the change of codon 56 from AAG to GAG was the only mutation in the studied individuals found within the cDNA segment coding for the abnormally migrating fragment of cdb3, we conclude that it represents the underlying molecular basis of the b3M polymorphism. We further support this conclusion by showing that electrophoresis in the presence of 4 mol/L urea abolished the difference in migration between proteolytic products of b3M and normal band 3, and that a fusion protein prepared from cDNA coding for the b3M allele again exhibits reduced electrophoretic mobility compared with the normal fusion protein. Finally, since most of the previously cloned mouse, rat, and chicken band 3 and band 3-related proteins contain glutamic acid in the position corresponding to amino acid 56 in the human band 3, we propose that the Memphis variant is the evolutionarily older form of band 3.

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