The Experts below are selected from a list of 156 Experts worldwide ranked by ideXlab platform
Elwira Lisowska - One of the best experts on this subject based on the ideXlab platform.
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The structures of Glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand
Glycobiology, 2014Co-Authors: David J. Ashline, Maria Duk, Elwira Lisowska, Jolanta Lukasiewicz, Vernon N. Reinhold, Ewa JaskiewiczAbstract:Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human Glycophorins - an update. Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte Glycophorin. Proc Natl Acad Sci USA, 72:2964-2968.) are well characterized but the structure of GPC N-glycans has remained unknown. This problem became important since it was reported that GPC N-glycans play an essential role in the interaction with Plasmodium falciparum EBA-140 merozoite ligand. The elucidation of these structures seems essential for full characterization of the GPC binding site for the EBA-140 ligand. We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in Glycophorin A N-glycans. Our results also open a possibility of new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC. We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure.
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Antigenic properties of human Glycophorins--an update.
Advances in experimental medicine and biology, 2001Co-Authors: Elwira LisowskaAbstract:Glycophorins are complex heavily glycosylated antigens carrying peptidic and glycopeptidic epitopes. Detailed immunochemical studies showed that GP AlGPB and GPC/GPD molecules have defined sites which are particularly immunogenic. These sites include N-terminal portions of all Glycophorins, internal fragments of their extracellular domains, and cytoplasmic tails. The extracellular epitopes involve directly oligosaccharide chains (e.g. blood group M- and N-related epitopes, or N-terminal epitopes of GPC) or have peptidic character, shown by the reaction of respective antibodies with synthetic peptides. Peptidic eitopes are independent of glycosylation, or are variably affected by adjacent 0- glycans which may mask the epitopes or may be required for a proper exposure of an antibody binding site. Several low incidence epitopes are present on variant Glycophorin molecules. Among anti-Glycophorin antibodies there are the ‘bispecific’ ones, or antibodies recognizing an epitope formed by an interaction of two proteins (Wrb). Alltogether, the Glycophorins serve as convenient model antigens for studying Ag-Ab interaction and a role of O-glycosylation in protein antigenic properties. Moreover, well defined specificty of monoclonal anti-Glycophorin antibodies makes them more precise tools in serological investigation and identification of normal and variant antigens. Last but not least, elucidation of antigenic properties of Glycophorins is important for identification and characterization of human anti-Glycophorin antibodies, which in some cases create medical problems at transfusion or pregnancy.
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Isolation and characterization of Glycophorin from nucleated (chicken) erythrocytes.
Archives of biochemistry and biophysics, 2000Co-Authors: Maria Duk, Hubert Krotkiewski, Taras Stasyk, Maxim Lutsik-kordovsky, Danuta Syper, Elwira LisowskaAbstract:Abstract A sialoglycoprotein fraction was isolated from chicken erythrocytes by two methods based on the phenol extraction or chloroform/2-propanol extraction of differently prepared erythrocyte membranes. Both preparations gave in SDS–PAGE two major PAS-stained bands (GP2 and GP3), which migrated as 60- and 33-kDa species, respectively, compared to reference proteins, or as 44- and 23-kDa molecules, compared to human Glycophorins. Some less abundant slower migrating PAS-stained components, antigenically related to GP2 and GP3, also were detected. No evidence for the presence of antigenically distinct glycoproteins of leukosialin type was obtained. Interconversion in SDS–PAGE, similar carbohydrate composition, and similar antigenic properties of GP2 and GP3 indicated that they are a dimer and monomer, respectively, of the same glycoprotein which shows properties that allow it to be classified as a Glycophorin. Lectin binding studies and methylation analysis of β-elimination products of chicken Glycophorin preparation showed the presence of O- glycans and N- glycans. The major O- glycans include sialylated Galβ1–3GalNAc units and more complex GlcNAc-containing chains. Among the N- glycans, there are complex-type biantennary structures with a bisecting GlcNAc residue, accompanied by chains with additional antennas linked to α-mannose residues. A characteristic feature of the chicken Glycophorin is a relatively high proportion of N- glycans to O- glycans, compared to the Glycophorin A from human erythrocytes.
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NOR polyagglutination and Sta Glycophorin in one family: relation of NOR polyagglutination to terminal α‐galactose residues and abnormal glycolipids
Transfusion, 1999Co-Authors: Grazyna Kuśnierz‐alejska, Marion E. Reid, Maria Duk, Jill R. Stony, Barbara Wiȩcek, Halina Seyfried, Elwira LisowskaAbstract:BACKGROUND: This report describes the characterization of polyagglutinable red cells (RBCs), identified in two generations of a Polish family. CASE REPORT: Untreated and modified RBCs of the proposita (TS) were tested by serologic methods, using human sera, antibodies, lectins, and inhibitors of agglutination. Moreover, Glycophorins were characterized by sodium docecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, and glycolipids were purified, fractionated by thin-layer chromatography, and detected with Ricinus communis agglutinin I (RCA-I, specific for galactose residues) and Griffonia simplicifolia IB4 lectin (GSL-IB4, specific for Galα1–3Gal- structure). Some of the experiments were also performed on RBCs of members of TS's family. RESULTS: Polyagglutination, found in four members of TS's family, was identified as the second case of an earlier described NOR polyagglutination. The polyagglutination was decreased by treating the RBCs with α-galactosidase and was inhibited by a neutral glycolipid fraction from NOR+ RBCs. Detection of neutral glycolipids of TS's RBCs on the thin-layer plate by RCA-I and GSL-IB4 revealed the presence of components that were not detectable in control RBCs. Moreover, Western blotting of RBC membranes from five family members with Glycophorin monoclonal antibodies and agglutination assays with anti-Sta and anti-Dantu sera identified the presence of Sta Glycophorin in four members of the family, two of whom were NOR+ and two NOR–. CONCLUSION: Our results showed that two rare features of TS's RBCs, NOR polyagglutination and Sta Glycophorin, are inherited independently, and that NOR+ RBCs contain neutral glycolipids with an abnormal oligosaccharide structure, most likely terminated with α-galactosyl residues.
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nor polyagglutination and sta Glycophorin in one family relation of nor polyagglutination to terminal α galactose residues and abnormal glycolipids
Transfusion, 1999Co-Authors: Grazyna Kuśnierzalejska, Marion E. Reid, Maria Duk, Jill R. Stony, Barbara Wiȩcek, Halina Seyfried, Elwira LisowskaAbstract:BACKGROUND: This report describes the characterization of polyagglutinable red cells (RBCs), identified in two generations of a Polish family. CASE REPORT: Untreated and modified RBCs of the proposita (TS) were tested by serologic methods, using human sera, antibodies, lectins, and inhibitors of agglutination. Moreover, Glycophorins were characterized by sodium docecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, and glycolipids were purified, fractionated by thin-layer chromatography, and detected with Ricinus communis agglutinin I (RCA-I, specific for galactose residues) and Griffonia simplicifolia IB4 lectin (GSL-IB4, specific for Galα1–3Gal- structure). Some of the experiments were also performed on RBCs of members of TS's family. RESULTS: Polyagglutination, found in four members of TS's family, was identified as the second case of an earlier described NOR polyagglutination. The polyagglutination was decreased by treating the RBCs with α-galactosidase and was inhibited by a neutral glycolipid fraction from NOR+ RBCs. Detection of neutral glycolipids of TS's RBCs on the thin-layer plate by RCA-I and GSL-IB4 revealed the presence of components that were not detectable in control RBCs. Moreover, Western blotting of RBC membranes from five family members with Glycophorin monoclonal antibodies and agglutination assays with anti-Sta and anti-Dantu sera identified the presence of Sta Glycophorin in four members of the family, two of whom were NOR+ and two NOR–. CONCLUSION: Our results showed that two rare features of TS's RBCs, NOR polyagglutination and Sta Glycophorin, are inherited independently, and that NOR+ RBCs contain neutral glycolipids with an abnormal oligosaccharide structure, most likely terminated with α-galactosyl residues.
Maria Duk - One of the best experts on this subject based on the ideXlab platform.
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The structures of Glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand
Glycobiology, 2014Co-Authors: David J. Ashline, Maria Duk, Elwira Lisowska, Jolanta Lukasiewicz, Vernon N. Reinhold, Ewa JaskiewiczAbstract:Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human Glycophorins - an update. Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte Glycophorin. Proc Natl Acad Sci USA, 72:2964-2968.) are well characterized but the structure of GPC N-glycans has remained unknown. This problem became important since it was reported that GPC N-glycans play an essential role in the interaction with Plasmodium falciparum EBA-140 merozoite ligand. The elucidation of these structures seems essential for full characterization of the GPC binding site for the EBA-140 ligand. We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in Glycophorin A N-glycans. Our results also open a possibility of new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC. We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure.
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Isolation and characterization of Glycophorin from nucleated (chicken) erythrocytes.
Archives of biochemistry and biophysics, 2000Co-Authors: Maria Duk, Hubert Krotkiewski, Taras Stasyk, Maxim Lutsik-kordovsky, Danuta Syper, Elwira LisowskaAbstract:Abstract A sialoglycoprotein fraction was isolated from chicken erythrocytes by two methods based on the phenol extraction or chloroform/2-propanol extraction of differently prepared erythrocyte membranes. Both preparations gave in SDS–PAGE two major PAS-stained bands (GP2 and GP3), which migrated as 60- and 33-kDa species, respectively, compared to reference proteins, or as 44- and 23-kDa molecules, compared to human Glycophorins. Some less abundant slower migrating PAS-stained components, antigenically related to GP2 and GP3, also were detected. No evidence for the presence of antigenically distinct glycoproteins of leukosialin type was obtained. Interconversion in SDS–PAGE, similar carbohydrate composition, and similar antigenic properties of GP2 and GP3 indicated that they are a dimer and monomer, respectively, of the same glycoprotein which shows properties that allow it to be classified as a Glycophorin. Lectin binding studies and methylation analysis of β-elimination products of chicken Glycophorin preparation showed the presence of O- glycans and N- glycans. The major O- glycans include sialylated Galβ1–3GalNAc units and more complex GlcNAc-containing chains. Among the N- glycans, there are complex-type biantennary structures with a bisecting GlcNAc residue, accompanied by chains with additional antennas linked to α-mannose residues. A characteristic feature of the chicken Glycophorin is a relatively high proportion of N- glycans to O- glycans, compared to the Glycophorin A from human erythrocytes.
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NOR polyagglutination and Sta Glycophorin in one family: relation of NOR polyagglutination to terminal α‐galactose residues and abnormal glycolipids
Transfusion, 1999Co-Authors: Grazyna Kuśnierz‐alejska, Marion E. Reid, Maria Duk, Jill R. Stony, Barbara Wiȩcek, Halina Seyfried, Elwira LisowskaAbstract:BACKGROUND: This report describes the characterization of polyagglutinable red cells (RBCs), identified in two generations of a Polish family. CASE REPORT: Untreated and modified RBCs of the proposita (TS) were tested by serologic methods, using human sera, antibodies, lectins, and inhibitors of agglutination. Moreover, Glycophorins were characterized by sodium docecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, and glycolipids were purified, fractionated by thin-layer chromatography, and detected with Ricinus communis agglutinin I (RCA-I, specific for galactose residues) and Griffonia simplicifolia IB4 lectin (GSL-IB4, specific for Galα1–3Gal- structure). Some of the experiments were also performed on RBCs of members of TS's family. RESULTS: Polyagglutination, found in four members of TS's family, was identified as the second case of an earlier described NOR polyagglutination. The polyagglutination was decreased by treating the RBCs with α-galactosidase and was inhibited by a neutral glycolipid fraction from NOR+ RBCs. Detection of neutral glycolipids of TS's RBCs on the thin-layer plate by RCA-I and GSL-IB4 revealed the presence of components that were not detectable in control RBCs. Moreover, Western blotting of RBC membranes from five family members with Glycophorin monoclonal antibodies and agglutination assays with anti-Sta and anti-Dantu sera identified the presence of Sta Glycophorin in four members of the family, two of whom were NOR+ and two NOR–. CONCLUSION: Our results showed that two rare features of TS's RBCs, NOR polyagglutination and Sta Glycophorin, are inherited independently, and that NOR+ RBCs contain neutral glycolipids with an abnormal oligosaccharide structure, most likely terminated with α-galactosyl residues.
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nor polyagglutination and sta Glycophorin in one family relation of nor polyagglutination to terminal α galactose residues and abnormal glycolipids
Transfusion, 1999Co-Authors: Grazyna Kuśnierzalejska, Marion E. Reid, Maria Duk, Jill R. Stony, Barbara Wiȩcek, Halina Seyfried, Elwira LisowskaAbstract:BACKGROUND: This report describes the characterization of polyagglutinable red cells (RBCs), identified in two generations of a Polish family. CASE REPORT: Untreated and modified RBCs of the proposita (TS) were tested by serologic methods, using human sera, antibodies, lectins, and inhibitors of agglutination. Moreover, Glycophorins were characterized by sodium docecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, and glycolipids were purified, fractionated by thin-layer chromatography, and detected with Ricinus communis agglutinin I (RCA-I, specific for galactose residues) and Griffonia simplicifolia IB4 lectin (GSL-IB4, specific for Galα1–3Gal- structure). Some of the experiments were also performed on RBCs of members of TS's family. RESULTS: Polyagglutination, found in four members of TS's family, was identified as the second case of an earlier described NOR polyagglutination. The polyagglutination was decreased by treating the RBCs with α-galactosidase and was inhibited by a neutral glycolipid fraction from NOR+ RBCs. Detection of neutral glycolipids of TS's RBCs on the thin-layer plate by RCA-I and GSL-IB4 revealed the presence of components that were not detectable in control RBCs. Moreover, Western blotting of RBC membranes from five family members with Glycophorin monoclonal antibodies and agglutination assays with anti-Sta and anti-Dantu sera identified the presence of Sta Glycophorin in four members of the family, two of whom were NOR+ and two NOR–. CONCLUSION: Our results showed that two rare features of TS's RBCs, NOR polyagglutination and Sta Glycophorin, are inherited independently, and that NOR+ RBCs contain neutral glycolipids with an abnormal oligosaccharide structure, most likely terminated with α-galactosyl residues.
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vicia villosa b4 lectin is the second anti tn lectin shown to react better with blood group n than m antigen
Glycoconjugate Journal, 1994Co-Authors: Maria Duk, Elwira LisowskaAbstract:Earlier studies showed thatMoluccella laevis lectin, which has anti-Tn specificity, reacts more strongly with native or desialylated blood group N Glycophorin A than with the respective Glycophorins of blood group M. We now present results indicating thatVicia villosa B4 anti-Tn lectin, which does not show detectable reaction with untreated Glycophorins or erythrocytes, reacts better with desialylated blood group N antigen than with asialo M antigen. This was demonstrated by three assays: (1) agglutination of asialoerythrocytes; (2) binding of biotinylated lectin to asialoerythrocytes immobilized on ELISA plates; and (3) inhibition of lectin binding to asialo-agalactoGlycophorin with asialoGlycophorins M and N. These results supply further support for the conclusion that Glycophorin of blood group N has more GalNAc residues unsubstituted with Gal (Tn receptors) than Glycophorin of blood group M.
John W Barnwell - One of the best experts on this subject based on the ideXlab platform.
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Plasmodium falciparum is able to invade erythrocytes through a trypsin-resistant pathway independent of Glycophorin B
Infection and immunity, 2003Co-Authors: Deepak Gaur, Marion E. Reid, John W Barnwell, Jill R. Storry, Louis H. MillerAbstract:Plasmodium falciparum invades erythrocytes through multiple ligand-receptor interactions, with redundancies in each pathway. One such alternate pathway is the trypsin-resistant pathway that enables P. falciparum to invade trypsin-treated erythrocytes. Previous studies have shown that this trypsin-resistant pathway is dependent on Glycophorin B, as P. falciparum strains invade trypsin-digested Glycophorin B-deficient erythrocytes at a highly reduced efficiency. Furthermore, in a recent study, the P. falciparum 7G8 strain did not invade Glycophorin B-deficient erythrocytes, a finding that was not confirmed in the present study. To analyze the degree of dependence on Glycophorin B for invasion by P. falciparum through the trypsin-resistant pathway, we have studied the invasion phenotypes of five parasite strains, 3D7, HB3, Dd2, 7G8, and Indochina I, on trypsin-treated normal and Glycophorin B-deficient erythrocytes. Invasion was variably reduced in Glycophorin B-deficient erythrocytes. Four strains, 3D7, HB3, Dd2, and Indochina I, invaded trypsin-treated erythrocytes, while invasion by the 7G8 strain was reduced by 90%. Among the four strains, invasion by 3D7, HB3, and Dd2 of trypsin-digested Glycophorin B-deficient erythrocytes was further reduced. However, Indochina I invaded trypsin-digested Glycophorin B-deficient erythrocytes at the same efficiency as its invasion of trypsin-digested normal erythrocytes. This strongly suggests that the Indochina I strain of P. falciparum is not dependent on Glycophorin B to invade through a trypsin-resistant pathway as are the strains 3D7, HB3, and Dd2. Thus, P. falciparum is able to invade erythrocytes through a Glycophorin B-independent, trypsin-resistant pathway.
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a plasmodium falciparum homologue of plasmodium vivax reticulocyte binding protein pvrbp1 defines a trypsin resistant erythrocyte invasion pathway
Journal of Experimental Medicine, 2001Co-Authors: Julian C Rayner, Esmeralda Vargasserrato, Curtis S Huber, Mary R Galinski, John W BarnwellAbstract:Invasion of erythrocytes by Plasmodium merozoites is an intricate process involving multiple receptor-ligand interactions. The Glycophorins and an unknown trypsin sensitive factor are all erythrocyte receptors used during invasion by the major human pathogen Plasmodium falciparum. However, only one erythrocyte receptor, Glycophorin A, has a well-established cognate parasite ligand, the merozoite protein erythrocyte binding antigen-175 (EBA-175). The involvement of several other parasite proteins during invasion have been proposed, but no direct evidence links them with a specific invasion pathway. Here we report the identification and characterization of P. falciparum normocyte binding protein 1 (PfNBP1), an ortholog of Plasmodium vivax reticulocyte binding protein-1. PfNBP1 binds to a sialic acid dependent trypsin-resistant receptor on the erythrocyte surface that appears to be distinct from known invasion receptors. Antibodies against PfNBP1 can inhibit invasion of trypsinized erythrocytes and two P. falciparum strains that express truncated PfNBP1 are unable to invade trypsinized erythrocytes. One of these strain, 7G8, also does not invade Glycophorin B–negative erythrocytes. PfNBP1 therefore defines a novel trypsin-resistant invasion pathway and adds a level of complexity to current models for P. falciparum erythrocyte invasion.
M J Tanner - One of the best experts on this subject based on the ideXlab platform.
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Localization of the protein 4.1-binding site on human erythrocyte Glycophorins C and D.
The Biochemical journal, 1994Co-Authors: N. J. Hemming, Me Reid, D J Anstee, W J Mawby, M J TannerAbstract:The flexibility of the human erythrocyte membrane is mediated by an underlying network of skeletal proteins which interact with the membrane through ankyrin and protein 4.1. The nature of the membrane attachment site(s) for protein 4.1 has yet to be fully elucidated. In this paper we show that purified protein 4.1 binds much less strongly to alkali-stripped membranes from erythrocytes of individuals with total Glycophorin C and D deficiency (Leach phenotype) than to alkali-stripped normal membranes. We further show that a synthetic peptide corresponding to amino acid residues 82-98 of the cytoplasmic domain of Glycophorin C specifically binds to purified protein 4.1 and inhibits protein 4.1 binding to alkali-stripped normal membranes. The same synthetic peptide binds directly to membranes from individuals with Glycophorin C and D deficiency but not to normal membranes. These results indicate that Glycophorins C and D provide major membrane attachment sites for protein 4.1 in normal erythrocytes and that this interaction is mediated by protein 4.1 binding to amino acid residues 82-98 of Glycophorin C and 61-77 of Glycophorin D.
Marion E. Reid - One of the best experts on this subject based on the ideXlab platform.
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Plasmodium falciparum is able to invade erythrocytes through a trypsin-resistant pathway independent of Glycophorin B
Infection and immunity, 2003Co-Authors: Deepak Gaur, Marion E. Reid, John W Barnwell, Jill R. Storry, Louis H. MillerAbstract:Plasmodium falciparum invades erythrocytes through multiple ligand-receptor interactions, with redundancies in each pathway. One such alternate pathway is the trypsin-resistant pathway that enables P. falciparum to invade trypsin-treated erythrocytes. Previous studies have shown that this trypsin-resistant pathway is dependent on Glycophorin B, as P. falciparum strains invade trypsin-digested Glycophorin B-deficient erythrocytes at a highly reduced efficiency. Furthermore, in a recent study, the P. falciparum 7G8 strain did not invade Glycophorin B-deficient erythrocytes, a finding that was not confirmed in the present study. To analyze the degree of dependence on Glycophorin B for invasion by P. falciparum through the trypsin-resistant pathway, we have studied the invasion phenotypes of five parasite strains, 3D7, HB3, Dd2, 7G8, and Indochina I, on trypsin-treated normal and Glycophorin B-deficient erythrocytes. Invasion was variably reduced in Glycophorin B-deficient erythrocytes. Four strains, 3D7, HB3, Dd2, and Indochina I, invaded trypsin-treated erythrocytes, while invasion by the 7G8 strain was reduced by 90%. Among the four strains, invasion by 3D7, HB3, and Dd2 of trypsin-digested Glycophorin B-deficient erythrocytes was further reduced. However, Indochina I invaded trypsin-digested Glycophorin B-deficient erythrocytes at the same efficiency as its invasion of trypsin-digested normal erythrocytes. This strongly suggests that the Indochina I strain of P. falciparum is not dependent on Glycophorin B to invade through a trypsin-resistant pathway as are the strains 3D7, HB3, and Dd2. Thus, P. falciparum is able to invade erythrocytes through a Glycophorin B-independent, trypsin-resistant pathway.
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NOR polyagglutination and Sta Glycophorin in one family: relation of NOR polyagglutination to terminal α‐galactose residues and abnormal glycolipids
Transfusion, 1999Co-Authors: Grazyna Kuśnierz‐alejska, Marion E. Reid, Maria Duk, Jill R. Stony, Barbara Wiȩcek, Halina Seyfried, Elwira LisowskaAbstract:BACKGROUND: This report describes the characterization of polyagglutinable red cells (RBCs), identified in two generations of a Polish family. CASE REPORT: Untreated and modified RBCs of the proposita (TS) were tested by serologic methods, using human sera, antibodies, lectins, and inhibitors of agglutination. Moreover, Glycophorins were characterized by sodium docecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, and glycolipids were purified, fractionated by thin-layer chromatography, and detected with Ricinus communis agglutinin I (RCA-I, specific for galactose residues) and Griffonia simplicifolia IB4 lectin (GSL-IB4, specific for Galα1–3Gal- structure). Some of the experiments were also performed on RBCs of members of TS's family. RESULTS: Polyagglutination, found in four members of TS's family, was identified as the second case of an earlier described NOR polyagglutination. The polyagglutination was decreased by treating the RBCs with α-galactosidase and was inhibited by a neutral glycolipid fraction from NOR+ RBCs. Detection of neutral glycolipids of TS's RBCs on the thin-layer plate by RCA-I and GSL-IB4 revealed the presence of components that were not detectable in control RBCs. Moreover, Western blotting of RBC membranes from five family members with Glycophorin monoclonal antibodies and agglutination assays with anti-Sta and anti-Dantu sera identified the presence of Sta Glycophorin in four members of the family, two of whom were NOR+ and two NOR–. CONCLUSION: Our results showed that two rare features of TS's RBCs, NOR polyagglutination and Sta Glycophorin, are inherited independently, and that NOR+ RBCs contain neutral glycolipids with an abnormal oligosaccharide structure, most likely terminated with α-galactosyl residues.
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nor polyagglutination and sta Glycophorin in one family relation of nor polyagglutination to terminal α galactose residues and abnormal glycolipids
Transfusion, 1999Co-Authors: Grazyna Kuśnierzalejska, Marion E. Reid, Maria Duk, Jill R. Stony, Barbara Wiȩcek, Halina Seyfried, Elwira LisowskaAbstract:BACKGROUND: This report describes the characterization of polyagglutinable red cells (RBCs), identified in two generations of a Polish family. CASE REPORT: Untreated and modified RBCs of the proposita (TS) were tested by serologic methods, using human sera, antibodies, lectins, and inhibitors of agglutination. Moreover, Glycophorins were characterized by sodium docecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, and glycolipids were purified, fractionated by thin-layer chromatography, and detected with Ricinus communis agglutinin I (RCA-I, specific for galactose residues) and Griffonia simplicifolia IB4 lectin (GSL-IB4, specific for Galα1–3Gal- structure). Some of the experiments were also performed on RBCs of members of TS's family. RESULTS: Polyagglutination, found in four members of TS's family, was identified as the second case of an earlier described NOR polyagglutination. The polyagglutination was decreased by treating the RBCs with α-galactosidase and was inhibited by a neutral glycolipid fraction from NOR+ RBCs. Detection of neutral glycolipids of TS's RBCs on the thin-layer plate by RCA-I and GSL-IB4 revealed the presence of components that were not detectable in control RBCs. Moreover, Western blotting of RBC membranes from five family members with Glycophorin monoclonal antibodies and agglutination assays with anti-Sta and anti-Dantu sera identified the presence of Sta Glycophorin in four members of the family, two of whom were NOR+ and two NOR–. CONCLUSION: Our results showed that two rare features of TS's RBCs, NOR polyagglutination and Sta Glycophorin, are inherited independently, and that NOR+ RBCs contain neutral glycolipids with an abnormal oligosaccharide structure, most likely terminated with α-galactosyl residues.
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The Glycophorin A Gene Family in Gorillas: Structure, Expression, and Comparison with the Human and Chimpanzee Homologues
Biochemical Genetics, 1997Co-Authors: S. S. Xie, Antoine Blancher, Marion E. Reid, C H Huang, Olga O. BlumenfeldAbstract:Homologues of MN blood group antigens, encoded by members of the Glycophorin A (GPA) gene family, are expressed in man, anthropoid apes, and some species of Old World monkeys. Previous studies had shown that a three-gene framework, most closely related to that in man, is present in the chimpanzee. Here we report the genomic structure, transcript map, and protein expression of the GYPA locus in gorillas. Compared to the corresponding human and chimpanzee homologues, gorilla GPA, GPB, and GPB/E genes each showed a high degree of sequence identity, with the same exon-intron organization. However, the expression of exons III, IV, or V encoding the extracellular or membrane domains of homologous Glycophorins varied among the three species. Gorilla GPA and GPB/E genes were unique in that the former occurred in two allelic forms with or without the expression of exon III, whereas the latter contained one (ψ exon III) instead of two silenced exons (ψ exons III and IV). Differences from human but not chimpanzee GPA also included the presence of a hybrid M/N epitope and the absence of the sequon for N-glycosylation. Owing to the retention of a functional exon III, gorilla GPB was more similar to chimpanzee GPB than human GPB. A transspecies allele was identified in the gorilla that gave rise to the Henshaw (He)-like antigen similar to that found in man. These results provide further insight into the model for evolution of the GPA gene family, indicating that the mechanisms underlying inter- and intraspecific polymorphism of Glycophorins could predate the divergence of gorillas as the consequence of gene duplication and diversification.