Recent studies have demonstrated that band 3 carries antigens of the Diego blood group system and have elucidated the molecular basis of several previously unassigned low incidence and high incidence antigens. Because the available serological data suggested that band 3 may carry additional low incidence blood group antigens, we screened band 3 genomic DNA encoding the membrane domain of band 3 for single-strand conformational polymorphisms. We found that the putative first ectoplasmic loop of band 3 carries blood group antigen ELO, 432 Arg→Trp; the third putative loop harbors antigens Vga (Van Vugt), 555 Tyr→His, BOW 561 Pro→Ser, Wu (Wulfsberg), 565 Gly→Ala, and Bpa (Bishop), 569 Asn→Lys; and the putative fourth ectoplasmic loop carries antigens Hga (Hughes), 656 Arg→Cys, and Moa (Moen), 656 Arg→His. We studied erythrocytes from carriers of five of these blood group antigens. We found similar levels of reticulocyte mRNA corresponding to the two band 3 gene alleles, normal content and glycosylation of band 3 in the red blood cell membrane, and normal band 3-mediated sulfate influx into red blood cells, suggesting that the mutations do not have major effect on band 3 structure and function. In addition to elucidating the molecular basis of seven low incidence blood group antigens, these results help to create a more accurate structural model of band 3.

BAND 3 (anion exchanger 1 [AE1]) is the most abundant integral protein of the red blood cell (RBC) membrane, being present in more than 1 million copies per RBC. It consists of two structurally and functionally largely independent domains. The N-terminal cytoplasmic domain of band 3 links the membrane to the underlying spectrin-based membrane skeleton and interacts with several glycolytic enzymes, hemoglobin, and hemichromes. The main function of the C-terminal membrane domain is to mediate exchange of chloride and bicarbonate anions across the plasma membrane. Current structural models predict that the membrane domain consists of 12 to 14 transmembrane helices connected by ectoplasmic and endoplasmic loops.1-4 The longest, fourth loop of band 3 is N-glycosylated1,4 and the carbohydrate chain carries more than half of the RBC ABO blood group epitopes.5 

Mutations of band 3 protein have been implicated in the pathogenesis of Southeast Asian ovalocytosis,6,7 hereditary spherocytosis,8-11 congenital acanthocytosis,12and, recently, distal renal tubular acidosis.13,14 An amino acid substitution in the cytoplasmic domain of band 3 underlies the abnormal electrophoretic mobility of a polymorphic band 3 variant known as band 3 Memphis.15,16 Yet another variant of band 3, known as band 3 Memphis II, displays, in addition to the abnormal electrophoretic mobility, an increased binding of the anion flux inhibitor dihydrodiisothiocyanatodisulfonate, disodium salt (H2DIDS).17 Spring et al18 reported that the Memphis II variant of band 3 carries the Dia blood group antigen.

Dia is a low incidence blood group antigen in Caucasians that is antithetical to Dib.19 Prevalence of Dia is much higher in American Indians, reaching up to 54% in some groups of South American Indians.20 The underlying polymorphism is a single amino acid substitution in position 854, with proline of the wild-type band 3 corresponding to the Dibantigen and leucine to the Dia antigen.21,22Dia and Dib became the first fully characterized antigens of the Diego blood group system assigned to the band 3 protein.21 Subsequently, Bruce et al23and our group24 have mapped the low incidence blood group antigen Wra to the C-terminal end of the fourth ectoplasmic loop. Glutamic acid 658 from the wild-type band 3 sequence underlies the high incidence antigen, Wrb, whereas the low incidence antigen, Wra, has lysine in the same position.

Recently, we and others have shown25,26 that three additional low incidence antigens, Rba,27WARR,28 and Wda,29 are associated with different single point mutations on band 3 and therefore belong to the Diego system. The amino acid changes associated with these three antigens are, respectively, 548Pro→Leu,30,31552Thr→Ile,32,33 and 557Val→Met.30,31,34,35 Although only one subject has been studied for another low incidence antigen, Tra, the experimental evidence strongly suggests that it is also located on the erythroid band 3 protein, with the underlying polymorphism being 551Lys→Asn.31 

Discovery of the molecular basis of these antigens yielded the first insight into the Diego blood group system as well as valuable information on the position and size of the external loops of band 3. In addition, in the case of Wrb, it helped to characterize the site of the band 3-glycophorin A interaction, because the Wrb antigen requires the presence of both these proteins for its expression.24,36,37 These results also suggested band 3 might carry additional low incidence antigens. We therefore reviewed the list of remaining low incidence antigens that had serological characteristics consistent with the possibility of their being carried by band 3 and that we had in liquid nitrogen storage. We found that antigens Bpa (Bishop),38 Wu (Wulfsberg),39 Moa (Moen),40Vga (Van Vugt),41 Hga(Hughes),42 BOW,43 and ELO44fulfilled these criteria and proceeded with characterization of their molecular basis.

Subjects.

Samples were obtained through the Serum, Cell and Rare Fluid (SCARF) exchange program or as gifts from numerous colleagues. Initial testing was performed on DNA isolated from blood samples stored in liquid nitrogen; additional testing was performed on freshly drawn blood samples shipped on ice to Boston. The subjects were heterozygotes for the studied blood group antigens.

DNA preparation.

DNA was isolated from blood samples (∼150 μL) that had been stored in liquid nitrogen or that had been obtained as fresh samples using the QIAamp Blood Kit (QIAGEN Inc, Chatsworth, CA). The manufacturer’s procedure for DNA isolation from whole blood was followed. The eluted DNA was used directly for polymerase chain reaction (PCR) amplification.

Single-strand conformational polymorphism (SSCP) analysis and sequencing of band 3 genomic DNA.

SSCP screening was performed according to Orita et al,45 46with minor modifications. Exons encoding the membrane domain of band 3 were PCR-amplified using pairs of intronic primers flanking the exons (Table 1; 35 cycles of 1 minute at 94°C and 1 minute at 60°C). To each 10 μL PCR reaction, 2.5 μCi32P-dATP (3,000 μCi/mmol; ICN, Costa Mesa, CA) was added. The PCR products were diluted in formamide loading buffer (86% formamide, 10% glycerol, 20 mmol/L EDTA, 0.25% bromophenol blue, 0.25% xylene cyanol), heat-denatured by boiling for 5 minutes, and quickly cooled on ice. Samples were electrophoresed for 16 hours at room temperature in a nondenaturing polyacrylamide mutation detection enhancement (MDE) gel (FMC BioProducts, Rockland, ME). Briefly, 25 mL MDE gel solution was mixed with 69 mL deionized water, 6 mL 10× TBE, 0.4 mL 10% ammonium persulfate, and 40 μL TEMED, and electrophoresis was performed at 8 W in the presence and at 4 W in the absence of 10% glycerol. The gel was exposed to Kodak XAR-5 film (Eastman Kodak, Rochester, NY) overnight at −80°C using intensifying screens. The band 3 gene exons displaying the SSCP were directly sequenced using the Sequenase version 2.0 DNA Sequencing Kit (Amersham, Arlington Heights, IL). We limited our SSCP screening to exons 11 to 20 of the band 3 gene, because these 10 exons encode the whole membrane domain of band 3 and mutations causing altered immunogenicity of band 3 are to be expected only in this portion of band 3.

Sequence comparisons and structural predictions.

All 12 currently known amino acid sequences of human,1,4mouse,2 rat,47 chicken,48,49 and trout50 erythroid band 3 proteins (AE1) and of the related anion exchangers AE2 from human,51,52 mouse,53rat,48 and rabbit54 and AE3 from human,55 mouse,56 and rat48 were retrieved from Genbank and aligned using the program CLUSTAL (PCGene; Intelligenetics, Mountain View, CA). Predictions of the number and position of band 3 transmembrane helices were made using programs RAOARGOS, HELIXMEM, and SOAP (PCGene) based on the reported human AE1 cDNA sequence.1 4 Based on these predictions, sequence comparisons, and available data on band 3 modifications by enzymes and chemicals with known cleavage and binding sites, we created a model of band 3 with 14 transmembrane helices.

Detection of band 3 mRNA alleles in reticulocyte RNA.

Total reticulocyte RNA was isolated by ammonium chloride lysis57 and reverse transcribed using random primers. cDNA was PCR-amplified using primers flanking the mutations, and the PCR product was digested with the appropriate restriction endonuclease. The digested PCR product was visualized by electrophoresis in agarose gels stained by ethidium bromide and the amount of DNA of individual bands was quantitated by densitometric scanning using the Eagle Eye II system (Stratagene, La Jolla, CA) and the ONE-Dscan 1.0 program (Scanalytics, Billerica, MA).

Sulfate fluxes and di-isothiocyano-dihydrostilbene disulphonate (DIDS) titration curves.

Cells were washed three times at 4°C in 140 mmol/L NaCl, 10 mmol/L Na phosphate, pH 7.4 (PBS), and three times in 84 mmol/L trisodium citrate, 1 mmol/L EGTA, pH 6.5, and subjected to assays of unidirectional disodium 35S-sulfate (ICN, Costa Mesa, CA) uptake in the absence or in the presence of increasing concentrations of the anion transport inhibitor DIDS (Molecular Probes, Eugene, OR), as described.10 Each flux study in RBCs carrying the studied antigens was performed in parallel using RBCs from unrelated subjects not carrying the antigens. The dependence of the sulfate influx on the increasing concentrations of DIDS was obtained using the linear least squares fit.

Enzyme treatment of intact RBCs.

One volume of washed RBCs was treated with 4 vol of α-chymotrypsin (5 mg/mL) for 1 hour at 37°C. Enzyme-treated cells were washed three times with PBS. Both treated and untreated antigen-positive RBCs were tested in parallel with the appropriate antisera.

Quantitation of erythrocyte membrane proteins.

Freshly drawn blood anticoagulated in acid citrate/dextrose was shipped on ice to Boston. Within 48 hours of phlebotomy, erythrocyte ghosts were prepared using the method of Dodge et al,58 with minor modifications described in Jarolim et al.8 Erythrocyte membrane proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by densitometry as described,9 and the relative amounts of RBC membrane proteins were expressed as ratios of integrated densitometric peaks of individuals’ proteins to the peak of band 3. The biochemical findings were correlated with the clinical data and the results were statistically evaluated.

SSCP are detected in three exons.

We detected SSCP polymorphisms in exon 12 in the carrier of the ELO antigen; in exon 14 for Vga, BOW, Wu, and Bpa; and in exon 16 for Hga and Moa(Fig 1).

Fig. 1.

Detection of seven SSCPs. SSCP screening detected polymorphisms in exon 12 in genomic DNA from the ELO heterozygote, four polymorphisms in exon 14, and two polymorphisms in exon 16. Two normal bands (n) visible in wild-type (wt) homozygotes correspond to complementary strands of PCR-amplified DNA. Heterozygosity is reflected by the presence of an additional 1 to 2 bands.

Fig. 1.

Detection of seven SSCPs. SSCP screening detected polymorphisms in exon 12 in genomic DNA from the ELO heterozygote, four polymorphisms in exon 14, and two polymorphisms in exon 16. Two normal bands (n) visible in wild-type (wt) homozygotes correspond to complementary strands of PCR-amplified DNA. Heterozygosity is reflected by the presence of an additional 1 to 2 bands.

Close modal
Sequence analysis of genomic DNA shows seven amino acid substitutions.

We PCR-amplified and directly sequenced exons containing the SSCPs. The detected mutations are summarized in Table2. We have verified the presence of these mutations in additional unrelated carriers of these seven blood group antigens with the exception of Vga, for which only related individuals were available. All seven mutations either create or abrogate restriction sites (Table 2). In six cases, we used PCR amplification followed by restriction digestion with an appropriate restriction endonuclease to confirm the presence of the mutation in additional carriers of the blood group antigen. Because endonuclease Tth 2 is not commercially available, we directly sequenced genomic DNA from the additional Bp(a+) subject.

Position of the mutated amino acids in band 3.

Figure 2 schematically depicts the membrane domain of band 3. The seven newly characterized blood group antigens are shown in bold. Previously characterized blood group antigens residing on band 3 are also shown. According to this scheme, six of seven mutated amino acids are located in the putative first, third, and fourth ectoplasmic loops of band 3, whereas asparagine 569, which is mutated to lysine in Bp(a+) subjects, is located at the external end of the sixth transmembrane segment. Proline 561, mutated to serine in the BOW-positive subject, is also relatively highly conserved; however, the degree of evolutionary conservation in the region of the third external loop shown in Table 2 changes to some extent with variations in the alignment parameters.

Fig. 2.

Antigens of the Diego blood group system. Schematic representation of the membrane domain of band 3 based on the structural predictions described in the Materials and Methods and Results. Positions of all antigens of the Diego blood group system are indicated; the seven antigens described in this report are shown in bold.

Fig. 2.

Antigens of the Diego blood group system. Schematic representation of the membrane domain of band 3 based on the structural predictions described in the Materials and Methods and Results. Positions of all antigens of the Diego blood group system are indicated; the seven antigens described in this report are shown in bold.

Close modal
Evolutionary conservation of mutated amino acids.

We aligned the five known AE1 amino acid sequences of the erythroid band 3 homologues as well as all 12 available sequences from the AE gene family and evaluated the conservation of individual amino acids. The results show (Table 2) that most of the mutated amino acids are not conserved in evolution. The only exception again appears to be the Bpa antigen (569 Asn→Lys) that is conserved in all 12 aligned amino acid sequences. Consequently, mutation of Asn 569 would be most likely to affect band 3 structure and anion exchange function.

Effects of enzyme treatment on RBC agglutinability.

We have digested intact RBCs from the carriers of all seven studied blood group antigens by trypsin, α-chymotrypsin, pronase, and ficin. The results of testing untreated and enzyme-treated antigen-positive RBCs are shown in Table 3. At least two sera containing antibodies to the seven low incidence antigens were tested on two independent experiments. Whereas the results for Vga, BOW, Wu, Hga, and Moa are expected, the results with ELO and Bpa were not and will be discussed later.

Similar quantities of normal and mutant mRNA alleles are detected in reticulocyte RNA.

We isolated and reverse-transcribed total reticulocyte RNA and amplified the cDNA as described. The PCR products were digested with the appropriate restriction endonuclease and electrophoresed in an ethidium bromide-stained gel. Densitometric scanning of the gel allowed us to estimate the relative content of mRNA corresponding to the two band 3 alleles of the heterozygous subjects. Using this semiquantitative technique, we have not detected significant differences between the content of the two band 3 cDNA alleles, suggesting that the mutations affect neither the transcription of the band 3 gene alleles nor the subsequent RNA processing (data not shown).

The mutations do not affect band 3 expression and function.

SDS-PAGE electrophoresis showed a normal electrophoretic pattern of band 3 protein in the carriers of the blood group antigens, including similar profiles of band 3 glycosylation (not shown). Subsequent densitometric scanning detected normal content of band 3 and other RBC membrane proteins. Because band 3 is the principal anion exchange protein of the RBC membrane, we compared anion fluxes in erythrocytes from carriers of the studied blood group antigens with those in normal RBCs. We did not detect significant differences between heterozygotes for the seven studied antigens and control subjects. Results of the DIDS titration of sulfate influx in the cells expressing the ELO, Wu, and Hga antigens are shown in Fig 3.

Fig. 3.

Band 3-mediated influx of radiolabeled sulfate. Influx of radiolabeled sulfate into control cells and cells from carriers of the seven low incidence blood group antigens was measured as described. No differences between the control wild-type cells and cells expressing the blood group polymorphisms were detected. Results are shown for the ELO, Wu, and Hga antigens.

Fig. 3.

Band 3-mediated influx of radiolabeled sulfate. Influx of radiolabeled sulfate into control cells and cells from carriers of the seven low incidence blood group antigens was measured as described. No differences between the control wild-type cells and cells expressing the blood group polymorphisms were detected. Results are shown for the ELO, Wu, and Hga antigens.

Close modal

As of 1995, 37 low incidence antigens were recognized as being discrete genetic characteristics unassigned to a particular blood group system.59 Many of the antibodies to these low incidence antigens occur together in multispecific sera.60 Often these same sera contain antibodies to glycophorin A determinants or to Wra, whose antithetical antigen Wrb has been shown to result from the interaction between glycophorin A and band 3.23,24 36 Thus, we undertook a concerted effort to identify band 3 polymorphisms recognized by these multispecific sera.

This work has led to the identification of seven new band 3 mutations. All substitutions were studied in more than one subject and the molecular basis of some of them was evaluated by enzyme treatment of intact, antigen-carrying erythrocytes. Based on these results, the International Society for Blood Transfusion (ISBT)/Working Party on Blood Group Terminology assigned the antigens to the Diego blood group as 010008 (ELO), 010009 (Wu), 010010 (Bpa), 010011 (Moa), 010012 (Hga), 010013 (Vga), and 010015 (BOW).

The clinical significance of the seven characterized antigens and their corresponding antibodies is unclear. The antigens do not represent a major problem in blood transfusion, because the majority of donors will lack the antigen. With the exception of ELO,61 62 they have not been reported in association with a hemolytic disease of the newborn.

Because it is not clear whether amino acids in the immediate vicinity of the mutated amino acid residues are sufficient to form the epitope of the blood group antigen or whether the epitope depends on the conformation of other portions of band 3, we treated RBCs with enzymes known to modify external loops of band 3. α-Chymotrypsin cleaves band 3 at tyrosines 553 and 555 of the third ectoplasmic loop and, quite predictably, α-chymotrypsin treatment abrogated agglutination of the Vga-, Wu-, and BOW-positive cells with the corresponding antibodies. The agglutination of Hga- and Moa-positive cells was not affected, because their epitopes are not located in the third loop. The reactivity of both examples of anti-ELO was unaffected by trypsin or ficin treatment of antigen-positive RBCs. However, whereas one anti-ELO was unaffected by α-chymotrypsin or pronase, the other was nonreactive with RBCs treated with either of these enzymes. Although this has been observed previously,63 the reason has not been determined. Perhaps some anti-ELO antibodies recognize an epitope formed by the interaction of loop 1 of band 3 with another membrane component that is α-chymotrypsin or pronase sensitive (such as the third loop of band 3). Unexpectedly, neither example of anti-Bpa agglutinated antigen-positive RBCs that had been treated with any of the enzymes. Again, it is possible that the epitope recognized by anti-Bpa requires an interaction of the third loop of band 3 with another, enzyme-sensitive component.

Comparison of the 12 known AE1, AE2, and AE3 sequences predicts a relatively low degree of conservation for the ectoplasmic loops compared with the transmembrane segments. Accordingly, five mutations occur in poorly conserved amino acids, whereas one, asparagine 569, that is mutated to Lys in the Bp(a+) subjects is conserved in all 12 AE homologues. The high degree of evolutionary conservation of asparagine 569 and its position at the beginning of the sixth transmembrane segment in the band 3 model (Fig 2) suggested that its substitution by positively charged lysine may have major structural and functional consequences. However, we have observed neither morphological abnormalities of the Bp(a+) RBCs nor differences between DIDS titration of sulfate influx in the Bp(a+) and control RBCs. Proline 561, mutated in BOW, is conserved in AE1 and AE2; however, only 3 of 5 AE1 sequences contain proline in the corresponding position. With the exception of BOW, we have studied DIDS-inhibitable sulfate influx in fresh erythrocytes from carriers of the other band 3 mutations and found no effect on sulfate influx. Based on these results, we propose that the first, third, and fourth ectoplasmic loops are not involved in the regulation of band 3-mediated anion exchange.

The main contributions of this work, besides clarifying the molecular basis of seven blood group antigens, are twofold. First, the results strongly support extracellular localization of the mutated amino acids. Because available serological data suggest that additional low incidence blood group antigens may be carried by band 3, ongoing characterization of such antigens may further improve the structural model of band 3.

Second, some of the antigens are located in the regions that have been implicated in the adhesion of modified RBCs to vascular endothelium.64-71 Band 3 was hypothesized to function as a receptor during invasion of human erythrocytes by Plasmodium falciparum.64 Naturally occurring anti-band 3 autoantibodies were found to recognize modified band 3 protein on the surface of Plasmodium falciparum-infected erythrocytes.65 Cytoadherence-related neoantigens onPlasmodium falciparum-infected human erythrocytes, resulting from the exposure of normally cryptic regions of the band 3 protein,66 were placed into the third ectoplasmic loop of band 3.67,68 The antibodies in sera of individuals living in a malaria-endemic region recognize peptide motifs from the extracellular loops of band 3,69 and the immune response to these band 3 neoantigens is associated with low parasite density.70 Monoclonal antibodies that react with human band 3 residues 542-555 from the third external loop recognize different band 3 conformations in uninfected and Plasmodium falciparum-infected erythrocytes.71 The cytoadherence of Plasmodium falciparum could be blocked both with synthetic peptides corresponding to motifs from the third ectoplasmic loop of band 3.67 Erythrocytes from carriers of low incidence blood group antigens in ectoplasmic loops of band 3 may serve as a model for evaluation of the sequence requirements for adhesion of Plasmodium falciparum-infected erythrocytes to the vascular endothelium.

Kay72 assigned the senescent RBC antigen to the third ectoplasmic loop, specifically amino acids 538-554. Erythrocytes carrying polymorphisms in the external loops of band 3 will again be useful for evaluation of the role of band 3 in erythrocyte cell aging. It is interesting to note that multiple antibodies to these low prevalence antigens are often found concomitantly in sera that contain autoreactive erythrocyte antibodies as well as in sera from individuals who have not received any prior RBC stimulus.

Finally, in a recent communication, Thevenin et al73reported that synthetic peptides derived from the second and third ectoplasmic regions of band 3 completely inhibited adherence of sickle cells to an endothelial monolayer in a static assay. These results again suggested that the third external loop of band 3 plays a role in the sequence-specific adhesion of modified RBCs to the vascular endothelium.

The majority of the data on the adhesion of parasitized and sickled erythrocytes have been obtained from in vitro studies using antibodies and synthetic peptides. Both these experimental approaches are prone to artifacts. Large antibody molecules may interfere with numerous interactions upon binding to the RBC surface. Inhibition of interactions by synthetic peptides is also frequently nonspecific, and the peptides may mimic other yet unknown antigens with similar or identical amino acid sequences. Erythrocytes from donors with substitutions in the ectoplasmic loops of band 3 represent therefore a valuable system for testing of the role of this portion of band 3 in erythrocyte aging, in cytoadherence of malarial parasites, or in adhesion of parasitized and sickled RBCs to the endothelium.

The authors thank the many colleagues who, over many years, sent us samples of RBCs expressing low incidence antigens. We also thank the following people who recently responded to requests for blood samples from selected donors: Ranjan Malde from the National Blood Service (London and the South East), London, UK; Graham Rowe from the National Blood Service (Wales), Cardiff, Wales; Judy Martin from the American Red Cross (Badger Region), Madison, WI; and Marilyn Moulds from Gamma Biologicals, Inc (Houston, TX). We thank Dr Jiri Zavadil. Milena Pohlova from the Institute of Hematology and Blood Transfusion (Prague, Czech Republic) for help with DNA sequencing and for preparation of the figures.

Supported in part by a grant from the National Blood Foundation (P.J.), by Grant No. 4118-3 from the Grant Agency of the Ministry of Health, Czech Republic (P.J.), and by a National Institutes of Health Specialized Center of Research (SCOR) grant in Transfusion and Medicine (HL 54459; M.E.R.).

The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. section 1734 solely to indicate this fact.

1
Lux
SE
John
KM
Kopito
RR
Lodish
HF
Cloning and characterization of band 3, the human erythrocyte anion-exchange protein (AE1).
Proc Natl Acad Sci USA
86
1989
9089
2
Kopito
RR
Lodish
HF
Structure of the murine anion exchange protein.
J Cell Biochem
29
1985
1
3
Kopito
RR
Lodish
HF
Primary structure and transmembrane orientation of the murine anion exchange protein.
Nature
316
1985
234
4
Tanner
MJA
Martin
PG
High
S
The complete amino acid sequence of the human erythrocyte membrane anion-transport protein deduced from the cDNA.
Biochem J
256
1988
703
5
Fukuda
M
Fukuda
MN
Changes in red cell surface glycoproteins and carbohydrate structures during the development and differentiation of human erythroid cells.
J Supramol Struct
17
1981
324
6
Jarolim
P
Palek
J
Amato
D
Hassan
K
Sapak
P
Nurse
GT
Rubin
HL
Zhai
S
Sahr
KE
Liu
SC
Deletion in erythrocyte band 3 gene in malaria-resistant Southeast Asian ovalocytosis.
Proc Natl Acad Sci USA
88
1991
11022
7
Schofield
AE
Tanner
MJA
Pinder
JC
Clough
B
Bayley
PM
Nash
GB
Dluzewski
AR
Reardon
DM
Cox
TM
Wilson
RJM
Gratzer
WB
Basis of unique red cell membrane properties in hereditary ovalocytosis.
J Mol Biol
223
1992
949
8
Jarolim
P
Palek
J
Rubin
HL
Prchal
JT
Korsgren
C
Cohen
CM
Band 3 Tuscaloosa: Pro327→Arg327 substitution in the cytoplasmic domain of erythrocyte band 3 protein associated with spherocytic hemolytic anemia and partial deficiency of protein 4.2.
Blood
80
1992
523
9
Jarolim
P
Rubin
HL
Liu
S-C
Cho
MR
Brabec
V
Derick
LH
Yi
SJ
Saad
STO
Alper
S
Brugnara
C
Golan
DE
Palek
J
Duplication of 10 nucleotides in the erythroid band 3 (AE1) gene in a kindred with hereditary spherocytosis and band 3 protein deficiency (band 3PRAGUE).
J Clin Invest
93
1994
121
10
Jarolim
P
Rubin
HL
Brabec
V
Chrobak
L
Zolotarev
AS
Alper
SL
Brugnara
C
Wichterle
H
Palek
J
Mutations of conserved arginines in the membrane domain of erythroid band 3 protein lead to a decrease in membrane-associated band 3 and to the phenotype of hereditary spherocytosis.
Blood
85
1995
634
11
Rybicki
AC
Qiu
JJH
Musto
S
Rosen
NL
Nagel
RL
Schwartz
RS
Human erythrocyte protein 4.2 deficiency associated with hemolytic anemia and a homozygous 40glutamic acid→lysine substitution in the cytoplasmic domain of band 3 (band 3Montefiore).
Blood
81
1993
2155
12
Bruce
LJ
Kay
MMB
Lawrence
C
Tanner
MJA
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.
Biochem J
293
1993
317
13
Bruce
LJ
Cope
DL
Jones
GK
Schofield
AE
Burley
M
Povey
S
Unwin
RJ
Wrong
O
Tanner
MJA
Familial distal renal tubular acidosis is associated with mutations in the red cell anion exchanger (band 3, AE1) gene.
J Clin Invest
100
1997
1693
14
Jarolim
P
Shayakul
C
Prabakaran
D
Jiang
LW
Stuarttilley
A
Rubin
HL
Simova
S
Zavadil
J
Herrin
JT
Brouillette
J
Somers
MG
Seemanova
E
Brugnara
C
Guaywoodford
LM
Alper
SL
Autosomal dominant distal renal tubular acidosis is associated in three families with heterozygosity for the R589H mutation in the AE1 (band 3) Cl-/HCO3- exchanger.
J Biol Chem
273
1998
6380
15
Yannoukakos
D
Vasseur
C
Driancourt
C
Blouquit
Y
Delaunay
J
Wajcman
H
Bursaux
E
Human erythrocyte band 3 polymorphism (band 3 Memphis): Characterization of the structural modification (Lys56→Glu) by protein chemistry methods.
Blood
78
1991
1117
16
Jarolim
P
Rubin
HL
Zhai
S
Sahr
KE
Liu
SC
Mueller
TJ
Palek
J
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
80
1992
1592
17
Hsu
L
Morrison
M
A new variant of the anion transport protein in human erythrocytes.
Biochemistry
24
1985
3086
18
Spring
FA
Bruce
LJ
Anstee
DJ
Tanner
MJA
A red cell band 3 variant with altered stilbene disulphonate binding is associated with the Diego (Dia) blood group antigen.
Biochem J
288
1992
713
19
Thompson
PR
Childers
DM
Hatcher
DE
Anti-Dib. First and second examples.
Vox Sang
13
1967
314
20
Levine
P
Robinson
EA
Layrisse
M
Arends
T
Dominguez-Sisco
R
The Diego blood factor.
Nature
177
1956
40
21
Bruce
LJ
Anstee
DJ
Spring
FA
Tanner
MJA
Band 3 Memphis variant II. Altered stilbene disulfonate binding and the Diego (Dia) blood group antigen are associated with the human erythrocyte band 3 mutation Pro854→Leu.
J Biol Chem
269
1994
16155
22
Jarolim
P
Rubin
HL
Moulds
JM
Molecular characterization of the Diego blood group antigen.
Blood
84
1994
237a
(abstr, suppl 1)
23
Bruce
LJ
Ring
SM
Anstee
DJ
Reid
ME
Wilkinson
S
Tanner
MJA
Changes in the blood group Wright antigens are associated with a mutation at amino acid 658 in human erythrocyte band 3: A site of interaction between band 3 and glycophorin A under certain conditions.
Blood
85
1995
541
24
Jarolim
P
Moulds
JM
Molecular characterization of the Wright blood group antigens. Academy of Clinical Laboratory Physicians and Scientists, 30th Annual Meeting
1995
33
NY. Syracuse, NY, SUNY Health Science Center
Syracuse
(abstr)
25
Jarolim
P
Murray
J
Rubin
H
Smart
E
Zelinski
T
Moulds
JM
The low incidence antigens Wda, Rba, and WARR are located on band 3. Abstracts of the 24th Congress of the International Society of Blood Transfusion.
1996
73
ISBT
Makuhari, Japan
(abstr)
26
Bruce
LJ
Zelinski
T
Ridgwell
K
Tanner
MJA
The low-incidence blood group antigen, Wda, is associated with the substitution Val557→Met in human erythrocyte band 3 (AE1).
Vox Sang
71
1996
118
27
Contreras
M
Stebbing
B
Mallory
DM
Bare
J
Poole
J
Hammond
W
The Redelberger antigen Rba.
Vox Sang
35
1978
397
28
Coghlan
G
Crow
M
Spruell
P
Moulds
M
Zelinski
T
A ‘new’ low-incidence red cell antigen, WARR: Unique to native Americans.
Vox Sang
68
1995
187
29
Lewis
M
Kaita
H
A “new” low incidence “Hutterite” blood group antigen Waldner (Wda).
Am J Hum Genet
33
1981
418
30
Jarolim
P
Murray
JL
Rubin
HL
Smart
E
Moulds
JM
Wda and Rba blood group antigens are located in the third ectoplasmic loop of the erythroid band 3 protein.
Blood
86
1995
445a
(abstr, suppl 1)
31
Jarolim
P
Murray
JL
Rubin
HL
Smart
E
Moulds
JM
Blood group antigens Rba, Tra, and Wda are located in the third ectoplasmic loop of erythrocyte band 3 protein.
Transfusion
37
1997
607
32
Jarolim
P
Murray
J
Rubin
HL
Coghlan
G
Zelinski
T
A Thr552→Ile substitution in erythroid band 3 gives rise to the Warrior blood group antigen.
Transfusion
36
1996
49S
(abstr, suppl)
33
Jarolim
P
Murray
JL
Rubin
HL
Coghlan
G
Zelinski
T
A Thr552→Ile substitution in erythroid band 3 gives rise to the Warrior blood group antigen.
Transfusion
37
1997
398
34
Bruce
LJ
Tanner
MJA
Zelinski
T
The low incidence blood group antigen, Wda, is associated with the substitution Val557→Met in human erythrocyte band 3.
Transfusion
35
1995
52S
(abstr, suppl)
35
Bruce
LJ
Zelinski
T
Ridgwell
K
Tanner
MJA
The low-incidence blood group antigen, Wd(a), is associated with the substitution Val(557)→Met in human erythrocyte band 3 (AE1).
Vox Sang
71
1996
118
36
Telen
MJ
Chasis
JA
Relationship of the human erythrocyte Wrb antigen to an interaction between glycophorin A and band 3.
Blood
76
1990
842
37
Huang
CH
Reid
ME
Xie
SS
Blumenfeld
OO
Human red blood cell Wright antigens: A genetic and evolutionary perspective on glycophorin A-band 3 interaction.
Blood
87
1996
3942
38
Liotta
I
Purpura
M
Dawes
BJ
Giles
CM
Some data on the low frequency antigens Wra and Bpa.
Vox Sang
19
1970
540
39
Kornstad
L
Howell
P
Jorgensen
J
Heier Larsen
AM
Wadsworth
LD
The rare blood group antigen, Wu.
Vox Sang
31
1976
337
40
Kornstad
L
Brocteur
J
A new, rare blood group antigen, Moa (MOEN), in Transfusion Congress: American Association of Blood Banks XXV Annual Meeting and International Society of Blood Transfusion XIII International Congress.
1972
58
American Association of Blood Banks
Washington, DC
41
Young
S
Vga: A new low incidence red cell antigen.
Vox Sang
41
1981
48
42
Rowe
GP
Hammond
W
A new low-frequency antigen, Hga (Hughes).
Vox Sang
45
1983
316
43
Chaves
MA
Leak
MR
Poole
J
Giles
CM
A new low-frequency antigen BOW (Bowyer).
Vox Sang
55
1988
241
44
Coghlan
G
Green
C
Lubenko
A
Tippett
P
Zelinski
T
Low-incidence red cell antigen ELO (700.51): Evidence for exclusion from thirteen blood group systems.
Vox Sang
64
1993
240
45
Orita
M
Iwahana
H
Kanazawa
H
Hayashi
K
Sekiya
T
Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms.
Proc Natl Acad Sci USA
86
1989
2766
46
Orita
M
Suzuki
Y
Sekiya
T
Hayashi
K
Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction.
Genomics
5
1989
874
47
Kudrycki
KE
Shull
GE
Primary structure of the rat kidney band 3 anion exchange protein deduced from a cDNA.
J Biol Chem
264
1989
8185
48
Kudrycki
KE
Newman
PR
Shull
GE
cDNA cloning and tissue distribution of mRNAs for two proteins that are related to the band 3 Cl−/HCO−3 exchanger.
J Biol Chem
265
1990
462
49
Cox
JV
Lazarides
E
Alternative primary structures in the transmembrane domain of the chicken erythroid anionic transporter.
Mol Cell Biol
8
1988
1327
50
Hubner
S
Michel
F
Rudloff
V
Appelhans
H
Amino acid sequence of band-3 protein from rainbow trout erythrocytes derived from cDNA.
Biochem J
285
1992
17
51
Gehrig
H
Muller
W
Appelhans
H
Complete nucleotide sequence of band 3 related anion transport protein AE2 from human kidney.
Biochim Biophys Acta
1130
1992
326
52
Alper
SL
Kopito
RR
Libresco
SM
Lodish
HM
Cloning and characterization of a murine band 3-related cDNA from kidney and from a lymphoid cell line.
J Biol Chem
263
1988
17092
53
Lindsey
AE
Schneider
K
Simmons
DM
Baron
R
Lee
BS
Kopito
RR
Functional expression and subcellular localization of an anion exchanger cloned from choroid plexus.
Proc Natl Acad Sci USA
87
1990
5278
54
Chow
A
Dobbins
JW
Aronson
PS
Igarashi
P
cDNA cloning and localization of a band 3-related protein from ileum.
Am J Physiol
263
1992
G345
55
Yannoukakos
D
Stuart-Tilley
A
Fernandez
HA
Fey
P
Duyk
G
Alper
SL
Molecular cloning, expression, and chromosomal localization of two isoforms of the AE3 anion exchanger from human heart.
Circ Res
75
1994
603
56
Kopito
RR
Lee
BS
Simmons
DM
Lindsey
AE
Morgans
CW
Schneider
K
Regulation of intracellular pH by a neuronal homolog of the erythrocyte anion exchanger.
Cell
59
1989
927
57
Goosens
M
Kan
YW
DNA analysis in the diagnosis of hemoglobin disorders.
Methods Enzymol
76
1981
805
58
Dodge
JT
Mitchell
C
Hanahan
DJ
The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes.
Arch Biochem Biophys
100
1963
119
59
Daniels
GL
Anstee
DJ
Cartron
JP
Dahr
W
Issitt
PD
Jorgensen
J
Kornstad
L
Levene
C
Lomasfrancis
C
Lubenko
A
Mallory
D
Moulds
JJ
Okubo
Y
Overbeeke
M
Reid
ME
Rouger
P
Seidl
S
Sistonen
P
Wendel
S
Woodfield
G
Zelinski
T
Blood Group Terminology 1995—ISBT Working Party on Terminology for Red Cell Surface Antigens.
Vox Sang
69
1995
265
60
Daniels
G
Human Blood Groups.
1995
Blackwell Science
London, UK
61
Ford
DS
Stern
DA
Hawksworth
DN
Lubenko
A
Pope
JM
Chana
HS
Better
PJ
Haemolytic disease of the newborn probably due to anti-ELO, an antibody to a low frequency red cell antigen.
Vox Sang
62
1992
169
62
Better
PJ
Ford
DS
Frascarelli
A
Stern
DA
Confirmation of anti-ELO as a cause of haemolytic disease of the newborn.
Vox Sang
65
1993
70
63
Reid
M
Lomas-Francis
C
The Blood Group Antigen Factsbook.
1997
12
Academic
San Diego, CA
64
Okoye
VCN
Bennett
V
Plasmodium falciparum malaria: Band 3 as a possible receptor during invasion of human erythrocytes.
Science
227
1985
169
65
Winograd
E
Sherman
IW
Naturally occurring anti-band 3 autoantibodies recognize a high molecular weight protein on the surface of Plasmodium falciparum infected erythrocytes.
Biochem Biophys Res Commun
160
1989
1357
66
Crandall
I
Sherman
IW
Cytoadherence-related neoantigens on Plasmodium falciparum (human malaria)-infected human erythrocytes result from the exposure of normally cryptic regions of the band 3 protein.
Parasitology
108
1994
257
67
Crandall
I
Collins
WE
Gysin
J
Sherman
IW
Synthetic peptides based on motifs present in human band 3 protein inhibit cytoadherence/sequestration of the malaria parasite Plasmodium falciparum.
Proc Natl Acad Sci USA
90
1993
4703
68
Iqbal
J
Siddique
AB
Ahlborg
N
Perlmann
P
Berzins
K
Cytoadherence-related homologous motifs in Plasmodium falciparum antigen Pf155/RESA and erythrocyte band 3 protein.
Parasitology
110
1995
503
69
Crandall
I
Guthrie
N
Sherman
IW
Plasmodium falciparum: Sera of individuals living in a malaria-endemic region recognize peptide motifs of the human erythrocyte anion transport protein.
Am J Trop Med Hyg
52
1995
450
70
Hogh
B
Petersen
E
Crandall
I
Gottschau
A
Sherman
IW
Immune response to band 3 neoantigens on Plasmodium falciparum-infected erythrocytes in subjects living in an area of intense malaria transmission are associated with low parasite density and high hematocrit value.
Infect Immun
62
1994
4362
71
Guthrie
N
Crandall
IE
Marini
S
Fasciglione
GF
Sherman
IW
Monoclonal antibodies that react with human band 3 residues 542-555 recognize different conformations of this protein in uninfected and Plasmodium falciparum infected erythrocytes.
Mol Cell Biochem
144
1995
117
72
Kay
MMB
Generation of senescent cell antigen on old cells initiates IgG binding to a neoantigen.
Cell Mol Biol
39
1993
131
73
Thevenin
BJM
Crandall
I
Ballas
SK
Sherman
IW
Shohet
SB
Band 3 peptides block the adherence of sickle cells to endothelial cells in vitro.
Blood
90
1997
4172

Author notes

Address reprint requests to P. Jarolim, MD, PhD, Department of Pathology, Brigham and Women’s Hospital, Harvard Medical School, 75 Francis St, Boston, MA 02111; e-mail: pjarolim@rics.bwh.harvard.edu.

Sign in via your Institution