Enzymatic Determination of the Anomeric Structures of Two Blood-Group B Active Glycosphingolipids Recombined with Apolipoproteins

Abstract Anomeric configuration of oligosaccharides usually is established by specific glycosidases. For this purpose detergents achieving water solubility of primarily insoluble glycosphingolipids as substrates have been replaced by delipidated hum an serum high density lipoproteins. The new method, tested by several well characterized glycosphingolipids and glycosidases, finally was applied to the evaluation of anomeric structures of two blood-group B active glycosphingolipids [ceramide hexa-saccharide (B-I) and ceramide octasaccharide (B -II)] from hum an erythrocyte membranes. In both B-I and B-II, α-glycosidic linkage was dem onstrated for the term inal galactose and fucose residues. β-glycosidic linkage has been evaluated for backbone saccharides. Together with the results pre viously obtained by composition analysis, linkage analysis and sequence analysis the following complete structure can be established:B -I: Galα1 → 3Gal (2 ← 1αFuc)β1 → 4GlcNAcβ1 → 3Galβ1 → 4Glcβ1 → 1Cer;B-II: Galα1 → 3Gal (2 ← 1αFuc)β1 → 4GlcNAcβ1 → 3Galβ1 → 4GlcNAcβ1 → 3Galβ1 → 4Glcβ1 → 1Cer.

Download Full-text

Comparative study of glycophorin A derived O-glycans from human Cad, Sd(a+) and Sd(a-) erythrocytes

Biochemical Journal ◽

10.1042/bj2320813 ◽

1985 ◽

Vol 232 (3) ◽

pp. 813-818 ◽

Cited By ~ 18

Author(s):

D Blanchard ◽

C Capon ◽

Y Leroy ◽

J P Cartron ◽

B Fournet

Keyword(s):

Blood Group ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Red Cells ◽

Erythrocyte Membranes ◽

Glycophorin A ◽

Methylation Analysis ◽

Dolichos Biflorus ◽

Major Acid

Glycophorin A was purified from the erythrocyte membranes of blood group Cad, Sd(a+) and Sd(a-) donors and the oligosaccharide alditols, obtained after alkaline borohydride degradation, separated by h.p.l.c. on an alkylamine silica gel column, were characterized by sugar analysis. Structure determination of the major acid components by methylation analysis, g.l.c.-m.s. and 1H-n.m.r. indicated that the three blood group Cad red cells under study (samples Cad., Bui. and Des.) carry the same pentasaccharide GalNAc(beta 1-4)[NeuAc(alpha 2-3)]Gal(beta 1-3)[NeuAc(alpha 2-6)]GalNAc -ol(Cad determinant) but in different amounts. This pentasaccharide, however, was absent from glycophorin A of Sd(a+) and Sd (a-) donors, suggesting that the Sda determinant is not associated with glycophorins. It was calculated that glycophorin A from the original Cad donor (Cad.) carries about 12 O-glycosidically linked pentasaccharide chains per molecule whereas only 2-3 of these chains were present in the samples from the two other unrelated Cad individuals (Bui. and Des.) It is well known from quantitative agglutination studies that the proportion of red cells which can be agglutinated by the Dolichos biflorus lectin varies from one Cad blood sample to another. Some are completely agglutinated (Cad. donor) whereas others are only partially agglutinated (Bui. and Des. donors) suggesting that some red cells might not carry the Cad determinants. From the results presented above and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis studies it is suggested that Cad red cells from Bui. and Des. do not carry a mixture of glycophorin A molecules with or without the Cad pentasaccharides but a spectrum of glycoprotein molecules with varying amounts of Cad determinants.

Download Full-text

Purification of the Human Blood Group B Gene-Associated 3-α-D-Galactosyltransferase by Biospecific Adsorption onto Group O Erythrocyte Membranes

Glycoconjugate Research ◽

10.1016/b978-0-12-301302-6.50088-9 ◽

1979 ◽

pp. 1019-1021 ◽

Cited By ~ 1

Author(s):

Leonard R. Carne ◽

Winifred M. Watkins

Keyword(s):

Blood Group ◽

Human Blood ◽

Erythrocyte Membranes ◽

Human Blood Group ◽

Group B

Download Full-text

Association of human erythrocyte membrane glycoproteins with blood-group Cad specificity

Biochemical Journal ◽

10.1042/bj2070497 ◽

1982 ◽

Vol 207 (3) ◽

pp. 497-504 ◽

Cited By ~ 27

Author(s):

J P Cartron ◽

D Blanchard

Keyword(s):

Blood Group ◽

Erythrocyte Membrane ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Membrane Receptors ◽

Erythrocyte Membranes ◽

Molar Ratio ◽

Membrane Glycoproteins ◽

Acetylneuraminic Acid ◽

Group B

Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of erythrocyte membranes from a blood-group-B individual with the rare Cad phenotype indicates a lower-than-normal mobility of the main sialoglycoproteins, suggesting an increase in apparent molecular mass of 3kDa and 2kDa respectively for glycoprotein alpha (synonym glycophorin A) and glycoprotein delta (synonym glycophorin B). Since the chief structural determinant of Cad specificity is N-acetylgalactosamine, the membrane receptors have been isolated by affinity binding on immobilized Dolichos biflorus (horse gram) lectin. The predominant species eluted from the gel was the abnormal glycoprotein alpha, whereas in control experiments no material could be recovered from the adsorbent incubated with group-B Cad-negative erythrocyte membranes. After partition of the membranes with organic solvents, the blood-group-Cad activity was found in aqueous phases containing the sialoglycoproteins, but not in the organic phases containing simple or complex glycolipids, which, however, retained the blood-group-B activity. The carbohydrate composition of highly purified lipid-free glycoprotein alpha molecules prepared from Cad and control erythrocytes was determined. Interestingly the molar ratio of N-acetylneuraminic acid to N-acetylgalactosamine was equal to 2:1 in the case of controls and equal to 1:1 in the case of Cad erythrocytes. Taken together these results suggest that Cad specificity is defined by N-acetylgalactosamine residues carried by the alkali-labile oligosaccharide chains attached to the erythrocyte membrane sialo-glycoproteins.

Download Full-text

Synthesis of rhamnosylated arginine glycopeptides and determination of the glycosidic linkage in bacterial elongation factor P

Chemical Science ◽

10.1039/c6sc03847f ◽

2017 ◽

Vol 8 (3) ◽

pp. 2296-2302 ◽

Cited By ~ 11

Author(s):

Siyao Wang ◽

Leo Corcilius ◽

Phillip P. Sharp ◽

Andrei Rajkovic ◽

Michael Ibba ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Elongation Factor ◽

Glycosidic Linkage ◽

Anomeric Configuration ◽

Elongation Factor P

We describe the synthesis and incorporation of α- and β-configured rhamnosyl arginine cassettes into Pseudomonas aeruginosa elongation factor P-derived glycopeptides. These were used to unequivocally determine the native anomeric configuration of the rhamnose moiety in EF-P.

Download Full-text

Significance of raised immunoglobulin M levels in cord blood of small-for-gestational-age infants.

Archives of Disease in Childhood ◽

10.1136/adc.53.11.895 ◽

1978 ◽

Vol 53 (11) ◽

pp. 895-898 ◽

Cited By ~ 10

Author(s):

T G Matthews ◽

C O'Herlihy

Keyword(s):

Cord Blood ◽

Blood Group ◽

Gestational Age ◽

Small For Gestational Age ◽

Intrauterine Infection ◽

Immunoglobulin M ◽

Hammersmith Hospital ◽

Group B

Cord IgM values were determined in small-for-gestational-age infants born at Hammersmith Hospital during a 5 1/2-year period. 121 (12.5%) infants had levels more than 0.2 g/l; in 92 these were between 0.21 and 0.3 g/l. In only 18 (14.8%) was a level of 0.4 g/l exceeded, and 5 proved cases of intrauterine infection--rubella (2), syphilis (2), and toxoplasmosis (1)--were in this group. The factor most often associated with cord IgM more than 0.4 g/l was prolonged rupture of the membranes. There was an increased incidence of blood group B among the mothers, probably reflecting the greater number of nonCaucasian women giving birth to small-for-gestational-age infants. Determination of cord IgM did not help significantly indiagnosis.

Download Full-text