The interaction of wheat germ agglutinin with keratan from cornea and nasal cartilage

Carlsson, Hans Erik; Lōnngren, Jörgen; Goldstein, Irwin J.; Christner, James E.; Jourdian, George W.

doi:10.1016/0014-5793(76)80011-7

Cited by 24 publications

(7 citation statements)

References 25 publications

(13 reference statements)

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“…1) further supports their localization at a nonprotein target such as hyaluronan. In spite of a previous study reporting that WGA does not interact with hyaluronan (Carlsson et al 1976), our assumption was corroborated by a concentration-dependent inhibition of WGA binding using hyaluronan oligosaccharides. IC 50 values observed with hyaluronan oligosaccharides were several times higher than those observed with the monosaccharide GlcNAc.…”

Section: Discussioncontrasting

confidence: 67%

Lectin binding patterns reflect the phenotypic status of in vitro chondrocyte models

Toegel

Plattner

et al. 2009

In Vitro Cell.Dev.Biol.-Animal

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In vitro studies using chondrocyte cell cultures have increased our understanding of cartilage physiology and the altered chondrocytic cell phenotype in joint diseases. Beside the use of primary cells isolated from cartilage specimens of donors, immortalized chondrocyte cell lines such as C-28/I2 and T/C-28a2 have facilitated reproducible and standardized experiments. Although carbohydrate structures appear of significance for cartilage function, the contribution of the chondrocyte glycocalyx to matrix assembly and alterations of the chondrocyte phenotype is poorly understood. Therefore, the present study aimed to evaluate the glycoprofile of primary human chondrocytes as well as of C-28/I2 and T/C-28a2 cells in culture. First, the chondrocytic phenotype of primary and immortalized cells was assessed using real-time reverse transcriptase polymerase chain reaction, immunofluorescence, and glycosaminoglycans staining. Then, a panel of lectins was selected to probe for a range of oligosaccharide sequences determining specific products of the O-glycosylation and N-glycosylation pathways. We found that differences in the molecular phenotype between primary chondrocytes and the immortalized chondrocyte cell models C-28/I2 and T/C-28a2 are reflected in the glycoprofile of the cells. In this regard, the glycocalyx of immortalized chondrocytes was characterized by reduced levels of high-mannose type and sialic acid-capped N-glycans as well as increased fucosylated O-glycosylation products. In summary, the present report emphasizes the glycophenotype as an integral part of the chondrocyte phenotype and points at a significant role of the glycophenotype in chondrocyte differentiation.

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Section: Discussioncontrasting

confidence: 67%

Lectin binding patterns reflect the phenotypic status of in vitro chondrocyte models

Toegel

Plattner

et al. 2009

In Vitro Cell.Dev.Biol.-Animal

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“…Although the most effective oligosaccharide inhibitors of this lectin are structures containing al ---> 2 bound fucose on galactose residues, structures with the a1 --* 3 linked fucose on N-acetylglucosamine residues are also known to interact with the lectin (32) . It is possible that the fucose also contributes to the WGA resistance, because substituents on C-3 of N-acetylglucosamine are known to block the interaction of this sugar with the lectin (33) .…”

Section: Discussionmentioning

confidence: 99%

Enzymatic basis for a lectin-resistant phenotype: increase in a fucosyltransferase in mouse melanoma cells.

Finne¹,

Burger²,

Prieels³

1982

The Journal of Cell Biology

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In the search for the biochemical basis of the control of glycosylation of cell surface carbohydrates, revertant clones were isolated from previously characterized wheat germ agglutinin-resistant clones of B16 mouse melanoma cells by selection for resistance to Lotus tetragonolobus lectin or to ricin. Comparison of the wheat germ agglutinin-resistant clones with the parent and revertant clones indicated that this phenotype was correlated with an increased sensitivity to the Lotus lectin, a 60-to 70-fold increase in a1 -* 3 fucosyltransferase activity and a decreased sialic acid content of the N-glycosidic chains of glycoproteins . The results suggest a novel type of control mechanism for lectin resistance, an increase in a glycosyltransferase activity . The presence of a1 ---> 3 bound fucose on N-acetylglucosamine residues would interfere with the addition of sialic acid by a2--> 3 linkages to galactose residues in the carbohydrate units, and this change could explain the resistance to wheat germ agglutinin and the increased sensitivity to the Lotus lectin . A change in a regulatory gene for the fucosyltransferase as a possible primary cause for the changed phenotype is discussed.The availability of lectin-resistant cell lines with altered carbohydrate moieties of cell surface glycoproteins and glycolipids has provided a powerful tool for the study of the biosynthesis and function of the complex carbohydrates in animal cells (1-4). Some of the variant cell lines appear to have a block at some level of the biosynthetic pathway of protein-bound oligosaccharides, but the enzymatic basis for the changed phenotype is usually not known . For the study of the biological role of glycoproteins and glycolipids, it would be important to know how the expression of cell surface carbohydrates is regulated.Many lectin-resistant variant cell lines differ from the parent lines with respect to more than one property (5-7). Because the occurrence of more than one genetic or epigenetic change even in single-step isolates cannot be ruled out, it is not known whether all the changes observed could be ascribed to one primary change. Other evidence should therefore be obtained by the isolation of independent variants or revenants (7,8) . The isolation of independent variants has so far been shown only for the lectin-resistant cells with a block in a specific Nacetylglucosaminyltransferase (9, 10), whereas the isolation of revenants has usually not been successful (1, 7) . We report here the isolation of variants and revenants that could be obtained repeatedly as well as their enzymic defect .Previous work from this laboratory has described a wheat THE JOURNAL OF CELL BIOLOGY " VOLUME 92 FEBRUARY 1982 277-282 © The Rockefeller University Press -0021-9525/82/02/0277/06 $1 .00 germ agglutinin (WGA)-resistant mouse melanoma cell line with many altered properties, including changes in cell adhesion and metastasis (11) . Analysis of the protein-bound carbohydrates revealed a structural change involving increased fucose-content and de...

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“…Our finding that pretreatment of sections with aryl sulphatase increased the reaction of STA and to a lesser extent WGA, throughout the trabecular meshwork, suggests that sulphation of the polysaccharide residues may be causing steric hindrance and inhibition of the binding of lectins, particularly WGA. The most likely glycoconjugates to be sulphated are the proteoglycans of which keratan sulphate, (which in bovine cornea is N-linked (Seno et al, 1965)) is the only one known to react with the lectins WGA and STA (Carlsson et al, 1976;Toda et al, 1981).…”

Section: Discussionmentioning

confidence: 99%

“…Carlsson et al (1976)found that WGA would precipitate bovine corneal keratan sulphate only after desulphation, whereas Toda et al (1981) observed that it bound WGA and STA even without desulphation, although the substitution of sulphate at C6 interfered with the binding of WGA more strongly than with STA. Biochemical studies have identified keratan sulphate among the glycosaminoglycans of human trabecular meshwork (Acott et al, 1985) and rabbit trabecular meshwork (Knepper et al, 1981), while human TM in organ culture has been shown to synthesize keratan sulphate (Acott et al, 1985).…”

Section: Discussionmentioning

confidence: 99%

Glycoconjugates of the human trabecular meshwork: a lectin histochemical study

et al. 1995

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Twelve specimens of resin-embedded human trabecular meshwork were probed with a panel of 21 biotinylated lectins, using an avidin-biotin peroxidase revealing system, in order to determine the normal pattern of saccharide expression in this tissue. High-mannose, intermediate and hybrid N-linked glycans, and complex N-linked bisected and non-bisected bi/tri-antennate glycans, as shown by the binding of Canavalia ensiformis (ConA), Pisum sativum (PSA), Lens culinaris (LCA) agglutinins and Phaseolus vulgaris erythroagglutinin (ePHA), were strongly expressed by the canal of Schlemm endothelium and juxtacanalicular tissue, but less so by the corneoscleal meshwork. Highly branced complex glycans were not found, as there was no binding by Phaseolus vulgaris leukoagglutinin (IPHA). Sialyl residues, especially those alpha 2,6-linked as demonstrated by strong Sambucus nigra (SNA) lectin staining, were also abundant in this area. N-acetyllactosamine sequences and some O-linked glycans were present in the trabecular meshwork, as shown by Solanum tuberosum (STA), Datura stramonium (DSA), and Jacalin (Jac) lectin binding, while fucose residues were not detected by Tetragonolobus purpureas (LTA) or Ulex europaeus-1 (UEA-1) agglutinins. These results indicate similarities with renal glomerular and vascular endothelium, although the lack of binding with UEA-1 agglutinin suggests differences which may relate to the specialized function of the trabecular meshwork. This study provides a baseline for comparative analysis of the glycans of human trabecular meshwork in pathological conditions such as primary open-angle glaucoma.

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The interaction of wheat germ agglutinin with keratan from cornea and nasal cartilage

Cited by 24 publications

References 25 publications

Lectin binding patterns reflect the phenotypic status of in vitro chondrocyte models

Lectin binding patterns reflect the phenotypic status of in vitro chondrocyte models

Enzymatic basis for a lectin-resistant phenotype: increase in a fucosyltransferase in mouse melanoma cells.

Glycoconjugates of the human trabecular meshwork: a lectin histochemical study

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