Antigenic Properties of Human Erythrocyte Glycophorins

Lisowska, Elwira

doi:10.1007/978-1-4613-1663-3_10

Cited by 36 publications

(23 citation statements)

References 135 publications

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“…The glycoprotein of l 15 kDa was also capable of forming aggregates in a similar molecular mass range. The tendency to form aggregates is a common phenomenon observed for membrane glycoproteins, as exemplified in the case of glycophorin [61].…”

Section: Discussionmentioning

confidence: 99%

α‐d‐Galactose‐bearing glycoproteins on the surface of stimulated murine peritoneal macrophages

Petryniak

Huard

Goldstein

1992

European Journal of Biochemistry

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Two glycoproteins were isolated from lysates of thioglycollate-stimulated, murine peritoneal macrophages by affinity chromatography on immobilized Gr@~bniu simplicifoliu I lectin and by preparative SDS/PAGE. The glycoproteins were readily labeled on the surface of intact macrophages with 3H and 1251. The labeled glycoproteins migrated as broad bands of molecular mass 92 -109 kDa and 11 5 -125 kDa. The mobility of the glycoproteins decreased only slightly after reduction with dithiothreitol, indicating the absence of intersubunit disulfide bridges. The 92-kDa and 11 5-kDa glycoproteins had p l 5.2 -5.4 and p l < 4, respectively. Digestion of both glycoproteins with agalactosidase released 23% oftheir 3H content and abolished their ability to bind to the G. simpliciJ?olia I lectin, showing that they contain terminal cc-D-galactosyl groups. After reduction with 2-mercaptoethanol, each glycoprotein fraction was sensitive to N-glycanase; the 11 5-kDa glycoproteins produced a smear with the front at approximately 67 kDa, whereas the 92-kDa glycoprotein gave two bands of 61 kDa and 75 kDa. Unreduced glycoproteins were insensitive to N-glycanase, suggesting the presence of intramolecular disulfide bonds. Although each glycoprotein fraction was sensitive to endoglycosidase H, this enzyme produced only slight changes in molecular mass when compared with N-glycanase. From these results as well as from the specificity of the enzymes involved, it is concluded that each glycoprotein fraction contains complex-type oligosaccharides and a small amount of highmannose and/or hybrid-type oligosaccharides. While each glycoprotein fraction was bound to Duturu strumonium lectin, they failed to react with anti-[i-(Den)] serum and their digestion with endo-Pgalactosidase did not cause a band shift in SDS/PAGE. Taken together, these results suggest the presence of N-acetyllactosamine units which are not arrayed in linear form but occur as single units, bound either to C2 and C6, or to C2 and C4, or both, of outer mannosyl residues on complex-type oligosaccharides. The glycoprotein(s) fraction precipitated with anti-[I (Step)] serum, suggesting the presence of branched lactosaminoglycans. Digestion of both glycoprotein fractions with a mixture of sialidase and 0-glycanase did not alter their mobility in SDSIPAGE, suggesting a lack or low content of 0-linked trisaccharides and tetrasaccharides. Each glycoprotein fraction was bound specifically to Samhucus nigra and Mauckiu umurensis immobilized lectins, indicating the presence of sialic acid linked x2,6 to subterminal D-galactose or N-acetylgalactosamine residues, and a2,3 to N-acetyllactosamine residues, respectively.

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Section: Discussionmentioning

confidence: 99%

α‐d‐Galactose‐bearing glycoproteins on the surface of stimulated murine peritoneal macrophages

Petryniak

Huard

Goldstein

1992

European Journal of Biochemistry

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“…Reference mAbs R18, 10,000 10,000 >20,000 >20,000 >20,000 >20,000 >20,000 >20,000 4 Values indicate the minimal concentration (pg/ml) of inhibitor needed for complete inhib ition of the agglutination. Glycophorins from a crude extract were modified at Glu residues using carbodiimide, at sialic acid residues using periodate, at Met residues using performic acid and at Lys and Arg residues using pentanedione.…”

Section: Identification Of Target Membrane Proteins On Immunoblotsmentioning

confidence: 99%

“…Several were reviewed by Lisowska [4] ; others have been produced more recently. Most of them are direct ed against the extramembranous domains and recognized epitopes either in the N-terminal regions of GPA and GPB, including blood group M-or N-specific mAbs [6][7][8][9][10][11][12], or epitopes within the residues 27-70 of GPA [10,[13][14][15][16][17][18].…”

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confidence: 99%

“…Only a few mAbs bind to the C-terminal cytoplas mic tail of GPA [20][21][22][23]. These antibodies were useful for the identification and characterization of hybrid molecules composed of the N-terminal portion of GPA and the C-ter minal portion of GPB (Lepore type hybrid) or, vice versa, of the N-and C-terminal portions of GPB and GPA, respec tively (anti-Lepore type hybrid) [1][2][3][4]. However, the value of antibodies as tools is increased when their epitopes are precisely defined and characterized; this has been done only for some of the many antiglycophorin antibodies.…”

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confidence: 99%

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Fine Characterization of a Series of New Monoclonal Antibodies Directed against Glycophorin A

Rasamoelisolo¹,

Czerwiński²,

Bruneau³

et al. 1997

Vox Sanguinis

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Objectives: Glycophorins A (GPA) and B (GPB) are the major sialoglycoproteins of the human erythrocyte (RBC) membrane. To prepare tools for the analysis of GPA and GPB, we produced a series of new monoclonal antibodies (mAbs) that identified epitopes of GPA. Methods: Seven murine monoclonal antibodies directed to glycophorin A (GPA) were fully characterized by agglutination of untreated and enzyme-treated human erythrocytes, inhibition of agglutination using chemically modified glycophorins and peptides from GPA, immunoblotting, and binding to synthetic peptides on plastic pins. Results: The antibodies identify epitopes located on four different portions of GPA: (1) NaM13-6D2 binds to the N-terminal portion of GPA and GPB carrying the N blood group antigen; (2) NaM26-3F4 recognizes the homologous portion of GPA and GPB corresponding to their amino acids 6-26; (3) NaM10-2H12, NaM16-IB10 and NaM10-6G4 are specific for the amino acid sequence 38-45 of GPA; and (4) NaM37-5F4 and NaM13-4E4 bind to the amino acid residues 119-124 located on the intracellular ponion of GPA. Conclusion: These antibodies represent precise tools to investigate GPA and related molecules in different cells and tissues.

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“…The significance of GPE for the antigen variants of the MNSs system remains to be elucidated (for review, see ref. [1][2][3][4][5][6][7]). …”

Section: Introductionmentioning

confidence: 99%

Miltenberger Subsystem of the MNSs Blood Group System

Dahr¹

1992

Vox Sanguinis

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The Miltenberger (Mi) classes represent a group of phenotypes for red cells that carry low frequency antigens associated with the MNSs blood group system. The antigens of this system are known to be located on two sialogly- coproteins denoted as glycophorin A (GP A) and GP B. The structural alterations of seven (classes I, II, III, V, VI, VII, Vili) Mi variants and a related variant (J. L.) have been elucidated. Based on these data and yet incomplete studies of the Mi antigens, the approximate structural alterations in class IV and IX may be predicted. In addition, knowledge of the various structures and partial characterization of the Mi antigens allows one to propose detailed hypotheses concerning the epitopes recognized by the various antibodies that define the Mi subsystem. The understanding of the Mi subsystem at the molecular level paves the way for future studies aimed at a more detailed elucidation of epitopes of Mi-related antibodies, the characterization of novel Mi variants and a search for hypothetical, hitherto unknown Mi-related antibodies.

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Antigenic Properties of Human Erythrocyte Glycophorins

Cited by 36 publications

References 135 publications

α‐d‐Galactose‐bearing glycoproteins on the surface of stimulated murine peritoneal macrophages

α‐d‐Galactose‐bearing glycoproteins on the surface of stimulated murine peritoneal macrophages

Fine Characterization of a Series of New Monoclonal Antibodies Directed against Glycophorin A

Miltenberger Subsystem of the MNSs Blood Group System

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