|
|||||||||
|
|
|
|
|
|
|
|
|
|
|
Prepublished online as a Blood First Edition Paper on September 5, 2002; DOI 10.1182/blood-2002-04-1285.
RED CELLS
From the Institut National de la Santé et de la
Recherche Médicale (INSERM) U76, Institut National de la
Transfusion Sanguine, Paris, France; INSERM U473, Service
d'Hématologie, I, Hopital Bicêtre, AP-HP, Faculté de
Medecine Paris-Sud, Le Kremlin Bicêtre, France; Center for
Host/Pathogen Interactions, University of California, San Francisco;
and The Jackson Laboratory, Bar Harbor, ME.
Rhnull red cells are characteristically
stomato-spherocytic. This and other evidence suggest that the Rh
complex represents a major attachment site between the membrane lipid
bilayer and the erythroid skeleton. As an attempt to identify the
linking protein(s) between the red cell skeleton and the Rh complex, we analyzed the expression of Rh, RhAG, CD47, LW, and glycophorin B
proteins in red cells from patients with hereditary spherocytosis associated with complete protein 4.2 deficiency but normal band 3 (4.2(-)HS). Flow cytometric and immunoblotting analysis
revealed a severe reduction of CD47 (up to 80%) and a slower mobility
of RhAG on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, possibly reflecting an overglycosylation state. Unexpectedly, 4.2 The Rh antigens are defined by the association of
membrane polypeptides which are missing from or severely reduced in red blood cells (RBCs) of rare Rhnull individuals who suffer a
clinical syndrome characterized by abnormalities of the red cell shape, cation transport, and membrane phospholipid organization (reviewed in
Cartron1 and Huang et al2). The core of this
complex is thought to be a tetramer composed of 2 Rh and 2 RhAG
subunits3 to which accessory chains (CD47, LW, and
glycophorin B [GPB]) are associated by noncovalent
linkages.4-7 Rh and RhAG proteins are encoded by
homologous genes and are erythroid specific.8,9 However,
homologs of RhAG (RhBG and RhCG or RhGK) are expressed in nonerythroid
tissues, thus defining a new RH gene
superfamily.10
Recent studies have shed new light on potential biologic properties and
functions of the Rh and Rh-related proteins.11 Rh proteins
exhibit significant homology with the NH In this study, we show that RBCs from 2 unrelated patients with
hereditary spherocytosis (HS) associated with a complete lack of
protein 4.2 due to distinct mutations in the EPB42 gene
(4.2(-)HS)27,28 exhibit a severe reduction of CD47 level
together with a modified electrophoretic pattern of RhAG. These results
suggest that protein 4.2, through interaction with CD47, is involved in
the linkage of the Rh complex to the red cell skeleton and/or in its
translocation to the cell membrane.
Blood samples
Antibodies
Human monoclonal antibodies (hMAbs) were as follows: anti-D (clone LOR15C9) was from A. Blancher (Toulouse, France); anti-Rhc (clone RaE11), anti-RhE (clone MCG8), and anti-K1 (clone T27S) were from the INTS; and anti-C was from Diagast (Bordeaux, France). Polyclonal antibodies (PAbs) were as follows: anti-Rhe was from the Etablissement de Transfusion Sanguine (Lyon, France); anti-s was from Oxytèle (Versailles, France); anti-S was from Bioatlantic; anti-Co3 (Sar. serum) was from the CNRGS; MPC8 raised against the C-terminal region of the Rh polypeptides was described earlier34; anti-p55 raised against synthetic peptide residues 25-47 of the human p55 cytoplasmic protein35 were from P. Bailly (INTS); anti-4.2 and anti-protein 4.1 were described elsewhere.27,36 The rat anti-murine CD47 MAb (clone miap301) was previously described.31 The rabbit anti-murine RhAG PAb was prepared by immunizing rabbits with a synthetic peptide (420ELDRNFFQHANHNHVEHEV438) corresponding to the carboxy terminal region of the mouse RhAG glycoprotein.8 Membrane preparations and Triton X-100 extractions Membranes from washed RBCs were prepared by hypotonic lysis.37 All fractions were prepared from red cells that had been deep-frozen under preservative medium, a procedure known to destroy reticulocytes. Accordingly, the initial reticulocyte counts would not interfere. Solubilization by nonionic detergent was performed by adding 150 µL of 1% Triton X-100 to 50 µL of packed ghosts in phosphate-buffered saline (PBS) and gentle shaking at 4°C for 15 minutes. Supernatants and pellets were separated by cenrifugation (20 000g for 30 minutes at 4°C). Pellets were resuspended in 200 µL of 1× Laemmli buffer. Equal volume of supernatant and pellet fractions were loaded on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE; 10% acrylamide) under nonreducing conditions.Western blot analysis For Western blot analysis, membrane proteins from whole ghost lysates or Triton X-100 soluble and insoluble fractions separated on SDS-PAGE (10% acrylamide) were transferred to filters and incubated with relevant primary antibodies. Following washing, the filters were incubated with the appropriate peroxidase-conjugated secondary antibody (anti-rabbit, anti-mouse, or anti-rat IgG) (Biosys, Compiègne, France). Immunoblots were visualized using the ECL chemiluminescent system (Amersham, Bucks, United Kingdom).Flow cytometric analysis Frozen blood cell samples, containing only mature RBCs from variant and control donors were resuspended in 50 µL PBS/0.2% (wt/vol) bovine serum albumin (BSA) and incubated for 30 minutes at 22°C with mMAbs or hMAbs, respectively. After several washes with PBS, the cells were incubated for 30 minutes at 22°C with fluorescein isothiocyanate (FITC)-conjugated F(ab')2 fragments of goat anti-mouse and of goat anti-human immunoglobulins (Immunotech, Marseille, France). When using anti-mouse antibodies, the cell surface antigen expression was quantified using calibration mouse IgG-coated beads (Qifikit; DAKO, Glostrup, Denmark) as standards, according to the manufacturer's instructions. The results were expressed as specific antibody binding capacity (SABC) units that proved to be directly proportional to the number of molecules bound per cell. When detected by human Mabs, antigen expression was estimated from the mean fluorescence intensity (MFI), given in arbitrary units.
Flow cytometric analysis of 4.2(-)HS RBCs Expression of the Rh complex on RBCs from the patients with the 4.2(-)Lisboa and 4.2(-)Nancy mutations was quantitatively estimated by flow cytometric analysis of Rh, RhAG, CD47, LW, and GPB expression level with specific MAbs and PAbs. Since the 4.2(-)Lisboa and 4.2(-)Nancy RBCs exhibited the DCcee and dccee phenotypes, respectively, normal RhD-positive and RhD-negative RBCs were used as controls. As shown in Table 1. this analysis revealed that CD47 expression was severely reduced (by 80%) in RBCs from both 4.2(-) patients, whereas Rh and RhAG were normally expressed. Anti-S and anti-s antibodies revealed that 4.2(-)Lisboa and 4.2(-)Nancy RBCs exhibited a normal expression level of GPB with the Ss and SS phenotypes, respectively. As expected, more LW was found on control RhD-positive than on RhD-negative RBCs, and the LW status of 4.2(-)Lisboa and 4.2(-)Nancy RBCs was in accordance with their RhD-positive and RhD-negative status, respectively.
The mean fluorescence intensity (MFI) values measured with human MAbs or PAbs raised against several blood group antigens or blood group-carrying proteins, including band 3, GPA, and GPC, that belong to the ankyrin-protein 4.2-band 3 and protein 4.1-p55-GPC complexes, respectively, did not reveal any deviation from normality in the 4.2(-) RBCs (Table 1). The only exception was CD44 whose expression was found to be 3-fold higher in the 4.2(-) RBCs, as compared with controls. Western blot analysis of 4.2(-)HS RBCs Accordingly to previous findings,27,28 we did not detect protein 4.2 in patients carrying the Lisboa and the Nancy mutations (Figure 1). The major 72-kDa protein 4.2 isoform was detected in equal amounts in RBC membrane proteins prepared from donors with the RhD-positive, RhD-negative, and Rhnull phenotypes (Figure 1A-B). As quantitative and qualitative controls, the p55 and 4.1 proteins, which are part of the GPC-4.1-p55 complex whose expression is independent of Rh, were detected with the same intensity in all samples. Then, the same blots were successively incubated with antibodies against Rh, RhAG, and CD47, the major members of the Rh complex. As positive and negative controls, anti-Rh, -RhAG, and -CD47 antibodies gave strong signals with membrane proteins from RhD-positive and RhD-negative individuals, but no staining or a very weak staining with RBC membranes from Rhnull individuals, as expected.1 In agreement with the flow cytometric analysis, a clear reduction of the amount of CD47 polypeptide and a normal level of Rh polypeptides were observed in both 4.2(-) RBC membrane preparations, using the B6H12 and MPC8 antibodies, respectively. While flow cytomeric analysis indicated normal expression of RhAG at the cell surface of 4.2(-) RBCs (see Table 1), careful examination of the immunostaining experiment with MAb 2D10 which recognizes the RhAG protein (45-70 kDa) revealed a slightly lower electrophoretic mobility of the RhAG component from the 4.2(-) samples as compared with controls.
Expression of CD47 and protein 4.2 in RBCs from variant Rh phenotypes Altered expression of CD47 was also detected in RBCs with some rare Rh variant phenotypes using several anti-CD47 MAbs (Table 2). As compared with RhD-positive and RhD-negative controls, RBCs from D-- and D.. phenotypes exhibited about 70% to 75% reduction in CD47 expression, a value similar to that observed with the 4.2(-) samples. A significant albeit less important decrease of CD47 was also observed in RBCs from Rh variants of the RN phenotype. However, CD47 expression was not altered in monocytes (750 000 ± 10 000 molecules/cell) and platelets (6000 ± 1000 molecules/cell) from these variants, as compared with controls (not shown). Western blot analysis confirmed that the CD47 expression was reduced in D-- and D.. RBCs to a level similar to that observed in the 4.2(-) samples (Figure 2A). As previously shown, CD47 was nearly undetectable in Rhnull membrane preparations.1 The protein 4.2 level and the electrophoretic migration of RhAG were not affected by the reduction of CD47 in D-- and Rhnull RBCs (Figure 2B).
Expression of Rh-like complex in RBCs from 4.2  / and CD47/ gene-targeted mice were
analyzed by Western blotting using antibodies raised against the murine
homologs of RhAG and CD47 and with the MPC8 anti-human Rh antibody
previously reported to cross-react with the mouse Rh
polypeptide.38 As shown in Figure
3, the 32-kDa and 45- to 75-kDa bands
corresponding to murine Rh and RhAG polypeptides, respectively, were
detected with the same intensity in all samples. Furthermore, the
electrophoretic mobility of the RhAG glycoprotein was not modified in
the gene-targeted samples as compared with controls. Immunostaining
with the anti-CD47 antibody illustrated the lack of CD47 in
CD47/ mice and revealed a normal level of CD47 in the
4.2/ sample. A normal CD47 expression in the absence of
protein 4.2 was confirmed by flow cytometric analysis of
4.2/ and control RBCs (MFI: 520 vs 540, respectively;
not shown). Finally, Ponceau or Coomassie blue stainings revealed a
normal protein 4.2 level in CD47/ RBCs and, as
expected, the lack of the 4.2-specific 72-kDa component in the
4.2/ sample.
Comparison of detergent-solubilized properties of Rh, RhAG, and
CD47 from normal and 4.2(-)HS human RBCs and from wild-type and
4.2
As in humans, the presence or absence of protein 4.2 in wild-type and
4.2
In this study, we provide the first evidence for an association between the Rh complex and the red cell membrane skeleton component protein 4.2. Our conclusion was based on flow cytometric and Western blot analysis of the expression of the Rh complex members Rh, RhAG, CD47, GPB, and LW in RBCs from 2 unrelated patients with complete 4.2 deficiency due to distinct mutations in the EPB42 gene. The rationale of this analysis was 2-fold: (1) integral membrane proteins Rh, RhAG, and CD47 interact with the membrane skeleton of erythroid cells20 (and references herein),22,23 and (2) Rh complex deficiency and protein 4.2 deficiency are both associated with HS.1,24,25 Indeed, the observations that both 4.1(-) and GPC(-) genetic conditions are associated with hereditary elliptocytosis (HE) and that GPC is sharply reduced in 4.1(-) RBCs provided the first evidence of the linkage between the integral protein GPC and the skeletal component protein 4.1, an association which has been experimentally confirmed.26 Thus, we hypothesized that the analysis of Rh complex expression in RBCs from HS patients with a well-defined primary defect of skeletal protein(s) might likewise provide critical information for the identification of the protein partner(s) that link Rh-associated proteins to the membrane skeleton. The major finding of this study was the observation of a severe
reduction of CD47 protein in both 4.2(-) HS patients. Conversely, expression level of the other members of the Rh complex (Rh, RhAG, LW,
and GPB) was normal. CD47 in 4.2(-)Lisboa and 4.2(-)Nancy RBCs,
estimated from flow cytometric analysis and confirmed by immunoblotting, was decreased by 80%. As a control, CD47 expression was not altered in 4.1(-) HE red cells (not shown). Bruce et
al39 discovered a new variant of protein 4.2 deficiency
and independently of our preliminary report40 also found
that CD47 is markedly reduced in the absence of protein 4.2. Of note,
we found that CD44 expression was 3-fold higher in the 4.2(-) HS RBCs,
as compared with controls. CD44 overexpression might reflect a
mechanism to compensate for the loss of CD47-mediated membrane-skeleton
anchoring, as postulated for the concomitant up-regulation of AE2 and
NHE1 and down-regulation of AE1 in
4.2 These observations suggest that protein 4.2 plays a major role in CD47 trafficking to the membrane or provides a major attachment site for the Rh complex via CD47. However, we found that the lack of protein 4.2 did not alter Triton X-100 extractability of Rh and RhAG proteins. While these experiments suggest that other component(s) are likely to be involved in the interaction of the Rh complex with the red cell skeleton, further studies will be necessary to determine to what extent protein 4.2 might modulate the strength of this attachment. A reduced electrophoretic mobility of RhAG on SDS-PAGE was also observed in both 4.2(-) samples, suggesting an overglycosylation of this glycoprotein. An overglycosylation state of RhAG was previously described in GPB-deficient RBCs42 and was correlated to a slowness of RhAG trafficking in the Golgi, suggesting that GPB plays a critical role in the translocation of RhAG to the plasma membrane. Therefore, we hypothesized that protein 4.2 and/or CD47 might be similarly associated with RhAG trafficking. However, as discussed below, analysis of Rh variants with CD47 deficiency ruled out such a role for CD47. A reduced expression of CD47 was previously found on RBCs from Rhnull and D-- variant phenotypes.1,43 We show here that these RBCs contain normal levels of protein 4.2. We further extend these observations to show that RBCs from donors with the D.. and RN variant Rh phenotypes also exhibit a reduced expression of CD47. That the (secondary) decrease of CD47, in the studied variants of the RH system, has no bearing on the amount of protein 4.2, whereas the (primary) absence of protein 4.2 caused a sharp reduction of CD47 may be accounted for by the fact that protein 4.2 is much more represented than CD47 (200 000 molecules/RBC vs 20 000-50 000 molecules/RBC) and is firmly bound to the cytoplasmic domain of band 3. Since D--, D.., and RN RBCs are characterized by the lack or decreased expression of the RhCcEe antigens, these results imply that CD47 expression is dependent on the RhCcEe polypeptide. Conversely, CD47 expression is not dependent on RhD polypeptide, as we show the same CD47 levels in RhD-positive and RhD-negative RBCs in the presence of protein 4.2. Importantly, the RHCE variants described above, despite the reduction of CD47, exhibit normal levels of RhAG and Rh polypeptides in contrast to Rhnull variants and a normal migration of RhAG on SDS-PAGE in contrast to 4.2(-) variants. Moreover, the RBC morphology is normal. These observations have 2 implications: (1) protein 4.2 but not CD47 might interfere with RhAG trafficking, and (2) the cell shape abnormalities typical of 4.2(-)HS and Rhnull RBCs do not result from the CD47 deficiency, per se, but from the lack or low expression of protein 4.2, and of Rh or RhAG, respectively. Hence, these observations strengthen the model in which Rh and RhAG define the core of the Rh complex, whereas CD47, together with GPB and LW, represent dispensable accessory chains. The observation that gene-targeted CD47 CD47 has recently been recognized as a marker of self on RBCs.
CD47-deficient mouse erythrocytes are rapidly cleared from the
bloodstream by splenic red pulp macrophages (that express CD47) when
transfused in wild-type (CD47+/+) recipients but not in
CD47 In order to investigate in an animal model the effect of a complete
lack of protein 4.2 on Rh complex expression, we analyzed RBCs from
gene-targeted 4.2 In conclusion, we provide evidence that cell surface expression of the Rh complex member CD47 depends on the skeletal protein 4.2. Since involment of the Rh complex in the linkage between the membrane lipid bilayer and the red cell skeleton is increasingly evident, we hypothesize that CD47 and protein 4.2 interact in RBCs. Additional studies will be necessary to determine (1) whether protein 4.2 is also involved in the membrane translocation of the Rh-related proteins, as suggested by the abnormal glycosylation of RhAG in the absence of protein 4.2, and (2) whether the skeleton association and the emerging transporter activity of Rh and RhAG might be modified by the absence of protein 4.2.
We thank all our colleagues who provided useful antibodies, Dr M. E. dos Santos (Hopital de Santo Antonio dos Capuchos, Lisboa, Portugal) and Dr O. Agulles (CTS, Nancy, France) for providing 4.2(-) blood samples.
Submitted April 30, 2002; accepted August 6, 2002.
Prepublished online as Blood First Edition Paper, September 5, 2002; DOI 10.1182/blood-2002-04-1285.
Supported in part by the Institut National de la Santé et de la Recherche Médicale (INSERM), the Centre National de la Recherche Scientifique (CNRS), the Institut National de Transfusion Sanguine (INTS), and National Institutes of Health grant HL64885 (L.L.P.).
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Presented in part at the 43rd Annual Meeting of the American Society of Hematology, Orlando, FL, December 7-11, 2001.46 Reprints: Yves Colin, INSERM U76, Institut National de Transfusion Sanguine, 6 rue Alexandre Cabanel, 75015 Paris, France; e-mail: colin{at}idf.inserm.fr.
1. Cartron JP. RH blood group system and molecular basis of Rh-deficiency. Baillieres Best Pract Res Clin Haematol. 1999;12:655-689[Medline] [Order article via Infotrieve]. 2. Huang CH, Liu PZ, Cheng JG. Molecular biology and genetics of the Rh blood group system. Semin Hematol. 2000;37:150-165[CrossRef][Medline] [Order article via Infotrieve].
3.
Eyers SA, Ridgwell K, Mawby WJ, Tanner MJ.
Topology and organization of human Rh (rhesus) blood group-related polypeptides.
J Biol Chem.
1994;269:6417-6423
4.
Agre P, Cartron JP.
Molecular biology of the Rh antigens.
Blood.
1991;78:551-563 5. Cartron JP, Agre P. Rh blood group antigens: protein and gene structure. Semin Hematol. 1993;30:193-208[Medline] [Order article via Infotrieve]. 6. Anstee DJ, Tanner MJ. Biochemical aspects of the blood group Rh (rhesus) antigens. Baillieres Clin Haematol. 1993;6:401-422[Medline] [Order article via Infotrieve]. 7. Cartron JP. Defining the Rh blood group antigens. Biochemistry and molecular genetics. Blood Rev. 1994;8:199-212[CrossRef][Medline] [Order article via Infotrieve]. 8. Kitano T, Sumiyama K, Shiroishi T, Saitou N. Conserved evolution of the Rh50 gene compared to its homologous Rh blood group gene. Biochem Biophys Res Commun. 1998;249:78-85[CrossRef][Medline] [Order article via Infotrieve]. 9. Matassi G, Cherif-Zahar B, Pesole G, Raynal V, Cartron JP. The members of the RH gene family (RH50 and RH30) followed different evolutionary pathways. J Mol Evol. 1999;48:151-159[CrossRef][Medline] [Order article via Infotrieve]. 10. Huang CH, Liu PZ. New insights into the Rh superfamily of genes and proteins in erythroid cells and nonerythroid tissues. Blood Cells Mol Dis. 2001;27:90-101[CrossRef][Medline] [Order article via Infotrieve]. 11. Cartron JP, Colin Y. Structural and functional diversity of blood group antigens. Transfus Clin Biol. 2001;8:163-199[CrossRef][Medline] [Order article via Infotrieve]. 12. Marini AM, Urrestarazu A, Beauwens R, Andre B. The Rh (rhesus) blood group polypeptides are related to NH4+ transporters. Trends Biochem Sci. 1997;22:460-461[CrossRef][Medline] [Order article via Infotrieve]. 13. Marini AM, Matassi G, Raynal V, Andre B, Cartron JP, Cherif-Zahar B. The human Rhesus-associated RhAG protein and a kidney homologue promote ammonium transport in yeast. Nat Genet. 2000;26:341-344[CrossRef][Medline] [Order article via Infotrieve].
14.
Westhoff CM, Ferreri-Jacobia M, Mak DO, Foskett JK.
Identification of the erythrocyte Rh blood group glycoprotein as a mammalian ammonium transporter.
J Biol Chem.
2002;277:12499-12502 15. Gahmberg CG, Karhi KK. Association of Rho(D) polypeptides with the membrane skeleton in Rho(D)-positive human red cells. J Immunol. 1984;133:334-337[Abstract]. 16. Ridgwell K, Tanner MJ, Anstee DJ. The Rhesus (D) polypeptide is linked to the human erythrocyte cytoskeleton. FEBS Lett. 1984;174:7-10[CrossRef][Medline] [Order article via Infotrieve]. 17. Paradis G, Bazin R, Lemieux R. Protective effect of the membrane skeleton on the immunologic reactivity of the human red cell Rho(D) antigen. J Immunol. 1986;137:240-244[Abstract].
18.
Bloy C, Blanchard D, Lambin P, et al.
Human monoclonal antibody against Rh(D) antigen: partial characterization of the Rh(D) polypeptide from human erythrocytes.
Blood.
1987;69:1491-1497 19. Bloy C, Blanchard D, Hermand P, Kordowicz M, Sonneborn HH, Cartron JP. Properties of the blood group LW glycoprotein and preliminary comparison with Rh proteins. Mol Immunol. 1989;26:1013-1019[CrossRef][Medline] [Order article via Infotrieve]. 20. Gane P, Le Van Kim C, Bony V, et al. Flow cytometry analysis of the association between blood group-related proteins and the detergent-insoluble material of K562 cells and erythroid precursors. Br J Haematol. 2001;113:1-10[CrossRef][Medline] [Order article via Infotrieve].
21.
Discher DE, Mohandas N, Evans EA.
Molecular maps of red cell deformation: hidden elasticity and in situ connectivity.
Science.
1994;266:1032-1035 22. Gimm A, Mouro I, Lambin P, Mohandas N, Cartron JP. Direct evidence for in situ interaction between Rh complex and membrane skeleton in intact red cells [abstract]. Vox Sang. 2000;78(suppl 1):14. 23. Dahl K, Westoff C, Discher D. Attachment of CD47 (IAP) to the erythrocyte cytoskeleton and visual colocalisation with the Rh complex [abstract]. Mol Cell Biol. 2001;12(suppl):1113. 24. Delaunay J. Genetic disorders of the red cell membrane. In: Jameson JL, ed. Principles of Molecular Medicine. Totowa NJ: Humana Press; 1998:191-195. 25. Tse WT, Lux SE. Red blood cell membrane disorders. Br J Haematol. 1999;104:2-13[CrossRef][Medline] [Order article via Infotrieve]. 26. Takakuwa Y. Regulation of red cell membrane protein interactions: implications for red cell function. Curr Opin Hematol. 2001;8:80-84[CrossRef][Medline] [Order article via Infotrieve].
27.
Hayette S, Dhermy D, dos Santos ME, et al.
A deletional frameshift mutation in protein 4.2 gene (allele 4.2 Lisboa) associated with hereditary hemolytic anemia.
Blood.
1995;85:250-256
28.
Beauchamp-Nicoud A, Morle L, Lutz HU, et al.
Heavy transfusions and presence of an anti-protein 4.2 antibody in 4. 2(-) hereditary spherocytosis (949delG).
Haematologica.
2000;85:19-24
29.
Cherif-Zahar B, Raynal V, D'Ambrosio AM, Cartron JP, Colin Y.
Molecular analysis of the structure and expression of the RH locus in individuals with D--, Dc-, and DCw- gene complexes.
Blood.
1994;84:4354-4360
30.
Rouillac C, Gane P, Cartron J, Le Pennec PY, Cartron JP, Colin Y.
Molecular basis of the altered antigenic expression of RhD in weak D(Du) and RhC/e in RN phenotypes.
Blood.
1996;87:4853-4861
31.
Lindberg FP, Bullard DC, Caver TE, Gresham HD, Beaudet AL, Brown EJ.
Decreased resistance to bacterial infection and granulocyte defects in IAP-deficient mice.
Science.
1996;274:795-798 32. Peters LL, Jindel HK, Gwynn B, et al. Mild spherocytosis and altered red cell ion transport in protein 4.2- null mice. J Clin Invest. 1999;103:1527-1537[Medline] [Order article via Infotrieve]. 33. Blanchard D, Tournamille C, Petit-Le Roux Y, et al. A new murine monoclonal antibody directed against the Fy6 antigen [abstract]. Vox Sang. 1998;74(suppl 1):71.
34.
Hermand P, Mouro I, Huet M, et al.
Immunochemical characterization of Rhesus proteins with antibodies raised against synthetic peptides.
Blood.
1993;82:669-676
35.
Ruff P, Speicher DW, Husain-Chishti A.
Molecular identification of a major palmitoylated erythrocyte membrane protein containing the src homology 3 motif.
Proc Natl Acad Sci U S A.
1991;88:6595-6599 36. Jons T, Drenckhahn D. Identification of the binding interface involved in linkage of cytoskeletal protein 4.1 to the erythrocyte anion exchanger. Embo J. 1992;11:2863-2867[Medline] [Order article via Infotrieve]. 37. Steck TL, Kant JA. Preparation of impermeable ghosts and inside-out vesicles from human erythrocyte membranes. Methods Enzymol. 1974;31:172-180[CrossRef][Medline] [Order article via Infotrieve]. 38. Apoil P, Blancher A. Sequences and evolution of mammalian RH gene transcripts and proteins. Immunogenetics. 1999;49:15-25[CrossRef][Medline] [Order article via Infotrieve]. 39. Bruce LJ, Ghosh S, King MJ, Layton M, Delaunay J, Tanner MJ. Severe deficiency of CD47 and reduction of RHAG associated with the absence of protein 4.2 in the red blood cell membrane [abstract]. Blood. 2001;98(suppl 1):10. 40. Colin Y, Mouro-Chanteloup I, Peters L, et al. Analysis of protein 4.2 deficient red cells suggests an interaction between the Rh complex members CD47 and RhCcEe and the skeletal protein 4.2 [abstract]. Blood. 2001;98(suppl 1):30. 41. Denker SP, Barber DL. Ion transport proteins anchor and regulate the cytoskeleton. Curr Opin Cell Biol. 2002;14:214-220[CrossRef][Medline] [Order article via Infotrieve].
42.
Ridgwell K, Eyers SA, Mawby WJ, Anstee DJ, Tanner MJ.
Studies on the glycoprotein associated with Rh (rhesus) blood group antigen expression in the human red blood cell membrane.
J Biol Chem.
1994;269:6410-6416 43. Poss MT, Swanson JL, Telen MJ, Lasky LC, Vallera DA. Monoclonal antibody recognizing a unique Rh-related specificity. Vox Sang. 1993;64:231-239[Medline] [Order article via Infotrieve].
44.
Oldenborg PA, Zheleznyak A, Fang YF, Lagenaur CF, Gresham HD, Lindberg FP.
Role of CD47 as a marker of self on red blood cells.
Science.
2000;288:2051-2054 45. Guerra-Shinohara EM, Barretto OC. The erythrocyte cytoskeleton protein 4.2 is not demonstrable in several mammalian species. Braz J Med Biol Res. 1999;32:683-687[Medline] [Order article via Infotrieve]. 46. Colin Y, Mouro-Chanteloup I, Peters L, Gane P, Le Van Kim C, Delaunay J, Cartron JP. Analysis of protein 4.2 deficient red cells suggests an interaction between the Rh complex members CD47 and RhCcEe and the skeletal protein 4.2 [abstract]. Blood. 2001;98:10a.
© 2003 by The American Society of Hematology.
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
|
 |
|
  S. E. Iismaa, B. M. Mearns, L. Lorand, and R. M. Graham Transglutaminases and Disease: Lessons From Genetically Engineered Mouse Models and Inherited Disorders Physiol Rev, July 1, 2009; 89(3): 991 - 1023. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. K. Tsai and D. E. Discher Inhibition of "self" engulfment through deactivation of myosin-II at the phagocytic synapse between human cells J. Cell Biol., March 5, 2008; 180(5): 989 - 1003. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Subramanian, E. T. Boder, and D. E. Discher Phylogenetic Divergence of CD47 Interactions with Human Signal Regulatory Protein {alpha} Reveals Locus of Species Specificity: IMPLICATIONS FOR THE BINDING SITE J. Biol. Chem., January 19, 2007; 282(3): 1805 - 1818. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Subramanian, R. Parthasarathy, S. Sen, E. T. Boder, and D. E. Discher Species- and cell type-specific interactions between CD47 and human SIRP{alpha} Blood, March 15, 2006; 107(6): 2548 - 2556. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. M. Toye, S. Ghosh, M. T. Young, G. K. Jones, R. B. Sessions, M. Ramauge, P. Leclerc, J. Basu, J. Delaunay, and M. J. A. Tanner Protein-4.2 association with band 3 (AE1, SLCA4) in Xenopus oocytes: effects of three natural protein-4.2 mutations associated with hemolytic anemia Blood, May 15, 2005; 105(10): 4088 - 4095. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Lopez, S. Metral, D. Eladari, S. Drevensek, P. Gane, R. Chambrey, V. Bennett, J.-P. Cartron, C. Le Van Kim, and Y. Colin The Ammonium Transporter RhBG: REQUIREMENT OF A TYROSINE-BASED SIGNAL AND ANKYRIN-G FOR BASOLATERAL TARGETING AND MEMBRANE ANCHORAGE IN POLARIZED KIDNEY EPITHELIAL CELLS J. Biol. Chem., March 4, 2005; 280(9): 8221 - 8228. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Ripoche, O. Bertrand, P. Gane, C. Birkenmeier, Y. Colin, and J.-P. Cartron Human Rhesus-associated glycoprotein mediates facilitated transport of NH3 into red blood cells PNAS, December 7, 2004; 101(49): 17222 - 17227. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. N. Dahl, R. Parthasarathy, C. M. Westhoff, D. M. Layton, and D. E. Discher Protein 4.2 is critical to CD47-membrane skeleton attachment in human red cells Blood, February 1, 2004; 103(3): 1131 - 1136. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Nicolas, C. Le Van Kim, P. Gane, C. Birkenmeier, J.-P. Cartron, Y. Colin, and I. Mouro-Chanteloup Rh-RhAG/Ankyrin-R, a New Interaction Site between the Membrane Bilayer and the Red Cell Skeleton, Is Impaired by Rhnull-associated Mutation J. Biol. Chem., July 3, 2003; 278(28): 25526 - 25533. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. N. Chernova, A. K. Stewart, L. Jiang, D. J. Friedman, Y. Z. Kunes, and S. L. Alper Structure-function relationships of AE2 regulation by Ca2+i-sensitive stimulators NH+4 and hypertonicity Am J Physiol Cell Physiol, May 1, 2003; 284(5): C1235 - C1246. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
 |
 |
|||||||
 |
 |
 |
 |
 |
 |
 |
 |
||
| Copyright © 2003 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
Top
Abstract
Introduction
), CD47-SIRP