Purification and characterization of a blood-group A reactive hemagglutinin from the snail Helix pomatia and a study of its combining site

Hammarström, Sten; Kabat, Elvin A.

doi:10.1021/bi00835a002

Cited by 275 publications

(104 citation statements)

References 37 publications

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“…Both lectins can precipitate blood group A substance, specifically agglutinate blood group A RBCs, and react with a-o-GalNAc-(1---~3) o-GalNAc, and o-GalNAc (12,14,15,28); these findings further support the suggestion that they share common receptors. However, DBA was found not to inhibit the binding of HPA on RBCs even at DBA'HPA ratios higher than 100:1; this suggests that HPA may have a much higher binding affinity for the primary receptors than DBA and that HPA may be able to bind to other sugar receptors which have little or no affinity for DBA.…”

supporting

confidence: 67%

“…A considerable number of lectins are blood group-specific, and some recognize A, B, H, Le b, and N determinants (9,14). In the present investigation, two blood group A-specific lectins, obtained from Helix pomatia (15)(16)(17)(18) and Dolichos biflorus (19)(20)(21), were used to react with genotype AO human RBCs, with the aim of studying the interaction of these molecules with their receptors on the cell surface membrane and elucidating this specific recognition process of cell-cell association and dissociation. …”

mentioning

confidence: 77%

“…HPA was also purified in E.A. Kabat's Laboratory, with polyleucyl hog A + H blood group substances as previously described (15). The purity of these lectin preparations is shown in the SDS PAGE pattern (Fig.…”

Section: Methodsmentioning

confidence: 99%

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Interaction of lectins with membrane receptors on erythrocyte surfaces.

Sung¹,

Kabat²,

Chien³

1985

The Journal of Cell Biology

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The interactions of human genotype AO erythrocytes (red blood cells) (RBCs) with N-acetylgalactosamine-reactive lectins isolated from Helix pomatia (HPA)and from Dolichos biflorus (DBA) were studied. Binding curves obtained with the use of tritium-labeled lectins showed that the maximal numbers of lectin molecules capable of binding to human genotype AO RBCs were 3.8 x 105 and 2.7 x 105 molecules/RBC for HPA and DBA, respectively. The binding of one type of lectin may influence the binding of another type. HPA was found to inhibit the binding of DBA, but not vice versa. The binding of HPA was weakly inhibited by a /~-D-galactose-reactive lectin isolated from Ricinus communis (designated RCA1). Limufus polyphemus lectin (LPA), with specificity for N-acetylneuraminic acid, did not influence the binding of HPA but enhanced the binding of DBA. About 80% of LPA receptors (N-acetylneuraminic acid) were removed from RBC surfaces by neuraminidase treatment. Neuraminidase treatment of RBCs resulted in increases of binding of both HPA and DBA, but through different mechanisms. An equal number (7.6 x 105) of new HPA sites were generated on genotypes AO and GO RBCs by neuraminidase treatment, and these new sites accounted for the enhancement (AO cells) and appearance (OO cells) of hemagglutinability by HPA. Neuraminidase treatment did not generate new DBA sites, but increased the DBA affinity for the existing receptors; as a result, genotype AO cells increased their hemagglutinability by DBA, while GO cells remained unagglutinable. The use of RBCs of different genotypes in binding assays with 3H-labeled lectins of known specificities provides an experimental system for studying cell-cell recognition and association.

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confidence: 67%

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confidence: 77%

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Interaction of lectins with membrane receptors on erythrocyte surfaces.

Sung¹,

Kabat²,

Chien³

1985

The Journal of Cell Biology

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“…Since it was observed that in certain cells, HPL interacts weakly with polymeric Nacetyl-D-glucosamine (20), the HPL-gold complex was preincubated with 3-100 mM N-acetyl-D-glucosamine, and further sugars such as N-acetylneuraminic acid, D-glucose, a-methyl-o-mannopyranoside, t .-fucose and ß-o-galactose . Another type of control consisted in the incubation of the sections first with an excess of native HPL (50 kg/ml) and second with the HPL-gold complex each for 30 min.…”

Section: Reagentsmentioning

confidence: 99%

“…To further substantiate such a notion, we applied a newly developed Cytochemical approach for the localization of sugar residues with lectins on thin sections of intact cells by electron microscopy (46,48) to intestinal goblet cells which synthesize large amounts of 0-linked glycoproteins (7,8,61), For this purpose the lectin from the snail Helix poinatia, which interacts specifically with terminal (nonreducing) a-N-acetyl-D-galactosamine residues (20,21,68), was bound to particles of colloidal gold and the complex used to determine the intracellular compartments to which this lectin binds. The observed labeling pattern strongly suggested that the Golgi apparatus is the initial site where attachment of GaINAc to proteins occurs.…”

mentioning

confidence: 99%

Cytochemical localization of terminal N-acetyl-D-galactosamine residues in cellular compartments of intestinal goblet cells: implications for the topology of O-glycosylation.

Roth¹

1984

The Journal of Cell Biology

216

101

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The O-linked oligosaccharides of mucin-type glycoproteins contain N-acetyl-Dgalactosamine (GaINAc) that is not found in N-linked glycoproteins . Because Helix pornatia lectin interacts with terminal GaINAc, we used this lectin, bound to particles of colloidal gold, to localize such sugar residues in subcellular compartments of intestinal goblet cells. When thin sections of low temperature Lowicryl K4M embedded duodenum or colon were incubated with Helix pomatia lectin-gold complexes, no labeling could be detected over the cisternal space of the nuclear envelope and the rough endoplasmic reticulum . A uniform labeling was observed over the first and several subsequent cis Golgi cisternae and over the last (duodenal goblet cells) or the two last (colonic goblet cells) trans Golgi cisternae as well as forming and mature mucin droplets. However, essentially no labeling was detected over several cisternae in the central (medial) region of the Golgi apparatus . The results strongly suggest that core Oglycosylation takes place in cis Golgi cisternae but not in the rough endoplasmic reticulum . The heterogenous labeling for GaINAc residues in the Golgi apparatus is taken as evidence that termination of certain O-oligosaccharide chains by GaINAc occurs in trans Golgi cisternae .Glycoproteins can be classified according to the nature of the linkage between the oligosaccharide chains and the polypeptide. The two main families are those having oligosaccharide chains linked N-glycosidically from N-acetyl-D-glucosamine to the amide nitrogen of asparagine and those possessing oligosaccharide chains O-glycosidically linked from N-acetyl-D-galactosamine (GaINAc)' to the hydroxyl groups of serine and/or threonine (22,28,35,36,72). The biosynthesis of Nlinked glycoproteins is currently considered to be a single pathway involving the assembly ofa lipid linked oligosaccharide (1, 41, 42), the en bloc transfer of the oligosaccharide from the lipid carrier to the nascent peptide chain (31, 32, 55), the processing of the oligosaccharide chains by glycosidases (12,18,24,27,38,(63)(64)(65)(66)) and the addition of terminal sugars by glycosyltransferases (5, 44, 57). The establishment of the carbohydrate to protein linkage in N-glycosylation seems to be a cotranslational process located in the rough endoplasmic reticulum and later steps of the assembly are thought to occur mainly in the Golgi apparatus (2,22,28). Much less information is available about the assembly of the THE JOURNAL OF CELL BIOLOGY " VOLUME 98 FEBRUARY 1984 399-406 0The Rockefeller University Press -0021-9525/84/02/0399/08 $1 .00 oligosaccharide chains of O-linked glycoproteins and its topology . The initial glycosylation reaction in the synthesis of O-linked glycoproteins involves the direct transfer ofGaINAc from uridine diphosphate (UDP)-GaINAc to the polypeptide by a UDP-GaINAc: polypeptide transferase and does not require oligosaccharide preassembly nor lipid intermediates (22,23,25,36,37,58,67). Opinions vary among investigators, however, regarding the su...

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