scholarly journals The carbohydrate moiety of band-3 glycoprotein of human erythrocyte membranes

1980 ◽  
Vol 187 (3) ◽  
pp. 677-686 ◽  
Author(s):  
T Tsuji ◽  
T Irimura ◽  
T Osawa
Keyword(s):  
Blood Group ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Band 3 ◽  
Carbohydrate Moiety ◽  
Erythrocyte Membranes ◽  
Gel Chromatography ◽  
Branching Points ◽  
Human Erythrocyte Membranes ◽  
Group A

Band-3 glycoprotein was purified from human blood-group-A erythrocyte membranes by selective solubilization and gel chromatography on Sepharose 6B in the presence of sodium dodecyl sulphate. The purified glycoprotein was subjected to hydrazinolysis in order to release the carbohydrate moiety. The released oligosaccharides were N-acetylated and applied to a column of DEAE-cellulose. Most of the band-3 oligosaccharides obtained were found to be free of sialic acids. When this neutral fraction was subjected to gel chromatography on a column of Sephadex G-50, two broad peaks were observed indicating that the band-3 glycoprotein was heterogeneous in the size of the oligosaccharide moieties. All fractions from gel chromatography were found to contain galactose, mannose, N-acetylglucosamine and fucose. The higher-molecular-weight (mol.wt. 3000-8000) peak consisted of fucose, mannose, galactose, N-acetylglucosamine and N-acetylgalactosamine in a molar proportion of 1.6:3.0:8.4:10.5:0.2. Most of these oligosaccharides were digested with a mixture of beta-galactosidase and beta-N-acetylhexosaminidase after alpha-L-fucosidase treatment to give a small oligosaccharide with the structure alpha Man2-beta Man-beta GlcNAc-GlcNAc. Methylation studies and limited degradation by nitrous acid deamination showed that the oligosaccharides contained the repeating disaccharide Gal beta 1→4GlcNAc beta 1→3, with branching points at C-6 of some of the galactose residues. These results indicate that a major portion of the band-3 oligosaccharide has a common core structure, with heterogeneity in the numbers of the repeating disaccharides, and contains fucose residues both in the peripheral portion and in the core portion. Haemagglutination tests were also carried out to determine the blood-group specificities of the glycoprotein and the results demonstrated the presence of both blood-group-H and I antigenic activities.

Solubilized Glycoprotein from Human Erythrocyte Membranes Possessing Blood Group A, B and H Activity

Vox Sanguinis ◽  
1969 ◽  
Vol 17 (4) ◽  
pp. 289-299 ◽  
Author(s):  
N. B. Whittemore ◽  
N. C. Trabold ◽  
C. F. Reed ◽  
R. I. Weed
Keyword(s):  
Blood Group ◽  
Human Erythrocyte ◽  
Erythrocyte Membranes ◽  
Blood Group A ◽  
Human Erythrocyte Membranes ◽  
Group A

scholarly journals A protein activator of Mg2+-dependent, Ca2+-stimulated ATPase in human erythrocyte membranes distinct from calmodulin

1980 ◽  
Vol 187 (2) ◽  
pp. 507-513 ◽  
Author(s):  
Douglas Mauldin ◽  
Basil D. Roufogalis
Keyword(s):  
Atpase Activity ◽  
Sucrose Gradient ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Erythrocyte Membranes ◽  
Dependent Manner ◽  
Calmodulin Binding ◽  
Heat Stable ◽  
Protein Activator ◽  
Human Erythrocyte Membranes

Treatment of extensively washed erythrocyte membranes with 0.1mm-EDTA decreased their Mg2+-dependent, Ca2+-stimulated ATPase [(Mg2++Ca2+)-ATPase] activity. An activator released by this treatment restored the (Mg2++Ca2+)-ATPase to its original value in a Ca2+-dependent manner. This activator was different from calmodulin, as determined by a number of criteria. It was retained on an Amicon XM-100 ultrafiltration membrane (molecular-weight cut-off 100000); it appeared in the void volume of Sephadex G-100 and G-75 columns; it was not retained on a DEAE-cellulose ion-exchange column at ionic strengths similar to those used to retain calmodulin; and it maximally activated (Mg2++Ca2+)-ATPase activity less than calmodulin and at a higher Ca2+ concentration. Like calmodulin, the activator is heat-stable. The activator fraction isolated on a 2.5–15% sucrose gradient in 0.16m-KCl showed a single band of mol.wt. 63000 and no calmodulin on 10%-polyacrylamide/sodium dodecyl sulphate gels. A trace amount of calmodulin was detected in the activator fraction by radioimmunoassay (approx. 10pg/ml of ‘ghosts’), but this amount was insufficient to account for the (Mg2++Ca2+)-ATPase activation. Furthermore, calmodulin-binding protein failed to inhibit (Mg2++Ca2+)-ATPase activity by more than 10–20% in the membrane preparations from which the activator was extracted. It was concluded that erythrocyte membranes contain a (Mg2++Ca2+)-ATPase activator that may attenuate the activation of the Ca2+-transport ATPase by calmodulin.

Solubilized Glycoprotein from Human Erythrocyte Membranes Possessing Blood Group A, B and H Activity

Vox Sanguinis ◽  
1969 ◽  
Vol 17 (4) ◽  
pp. 289-299
Author(s):  
N.B. Whittemore ◽  
N.C. Trabold ◽  
C.F. Reed ◽  
R.I. Weed
Keyword(s):  
Blood Group ◽  
Human Erythrocyte ◽  
Erythrocyte Membranes ◽  
Blood Group A ◽  
Human Erythrocyte Membranes ◽  
Group A

Blood group A determinants with mono- and difucosyl type 1 chain in human erythrocyte membranes

Biochemistry ◽  
1985 ◽  
Vol 24 (14) ◽  
pp. 3578-3586 ◽  
Author(s):  
Henrik Clausen ◽  
Steven B. Levery ◽  
John M. McKibbin ◽  
Senitiroh Hakomori
Keyword(s):  
Blood Group ◽  
Human Erythrocyte ◽  
Erythrocyte Membranes ◽  
Blood Group A ◽  
Human Erythrocyte Membranes ◽  
Group A

Further characterization of type 2 and type 3 chain blood group A glycosphinogolipids from human erythrocyte membranes

Biochemistry ◽  
1986 ◽  
Vol 25 (22) ◽  
pp. 7075-7085 ◽  
Author(s):  
Henrik Clausen ◽  
Steven B. Levery ◽  
Edward Nudelman ◽  
Michael Baldwin ◽  
Senitiroh Hakomori
Keyword(s):  
Blood Group ◽  
Human Erythrocyte ◽  
Erythrocyte Membranes ◽  
Blood Group A ◽  
Human Erythrocyte Membranes ◽  
Group A ◽  
Type 3

scholarly journals Ultrastructure of the intact skeleton of the human erythrocyte membrane.

1986 ◽  
Vol 102 (3) ◽  
pp. 997-1006 ◽  
Author(s):  
B W Shen ◽  
R Josephs ◽  
T L Steck
Keyword(s):  
Human Erythrocyte ◽  
Actin Filaments ◽  
Band 3 ◽  
Carbon Films ◽  
Erythrocyte Membranes ◽  
Accessory Proteins ◽  
Red Cell Membrane ◽  
Human Erythrocyte Membranes ◽  
Low Ionic Strength ◽  
Triton X

Filamentous skeletons were liberated from isolated human erythrocyte membranes in Triton X-100, spread on fenestrated carbon films, negatively stained, and viewed intact and unfixed in the transmission electron microscope. Two forms of the skeleton were examined: (a) basic skeletons, stripped of accessory proteins with 1.5 M NaCl so that they contain predominantly polypeptide bands 1, 2, 4.1, and 5; and (b) unstripped skeletons, which also bore accessory proteins such as ankyrin and band 3 and small plaques of residual lipid. Freshly prepared skeletons were highly condensed. Incubation at low ionic strength and in the presence of dithiothreitol for an hour or more caused an expansion of the skeletons, which greatly increased the visibility of their elements. The expansion may reflect the opening of spectrin from a compact to an elongated disposition. Expanded skeletons appeared to be organized as networks of short actin filaments joined by multiple (5-8) spectrin tetramers. In unstripped preparations, globular masses were observed near the centers of the spectrin filaments, probably corresponding to complexes of ankyrin with band 3 oligomers. Some of these globules linked pairs of spectrin filaments. Skeletons prepared with a minimum of perturbation had thickened actin protofilaments, presumably reflecting the presence of accessory proteins. The length of these actin filaments was highly uniform, averaging 33 +/- 5 nm. This is the length of nonmuscle tropomyosin. Since there is almost enough tropomyosin present to saturate the F-actin, our data support the hypothesis that tropomyosin may determine the length of actin protofilaments in the red cell membrane.

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