Carrier-immobilized derivatized lysoganglioside GM1 is a ligand for specific binding sites in various human tumor cell types and peripheral blood lymphocytes and monocytes

Gabius, Sigrun; Kayser, Klaus; Hellmann, Klaus P.; Ciesiolka, Thomas; Trittin, Andrea; Gabius, H.‐J.

doi:10.1016/0006-291x(90)91459-6

Cited by 25 publications

(10 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The detection of shifts should therefore only be regarded as semiquantitative evidence. In comparison to other methods for glycan chain conjugation, the degree of modification of albumin by lysogangliosides or high mannose-type glycopeptides and a homobifunctional cross-linker yielded a similar extent (17,36,37). Since already one sugar chain on a glycoprotein can govern certain aspects of its physiological behavior, as for example seen for plasma clearance (38), these results encourage comparative assays of the three neoglycoprotein preparations to delineate to what extent the structural changes will translate into different properties in the interaction with isolated sugar receptors and binding sites in cells.…”

Section: Resultsmentioning

confidence: 99%

Neoglycoproteins with the Synthetic Complex Biantennary Nonasaccharide or Its α2,3/α2,6-Sialylated Derivatives: Their Preparation, Assessment of Their Ligand Properties for Purified Lectins, for Tumor Cells in Vitro, and in Tissue Sections, and Their Biodistribution in Tumor-Bearing Mice

et al. 1997

Self Cite

View full text Add to dashboard Cite

Neoglycoproteins were prepared with chemoenzymatically synthesized complex biantennary N-glycan derivatives the nonreducing ends of which bear typical sequences found in glycoproteins. A chemically obtained biantennary heptasaccharide-azide was reduced and acylated with a 6-aminohexanoyl spacer. Elongation of the deprotected heptasaccharide using glycosyltransferases yielded a biantennary nonasaccharide with terminal galactose residues and two undecasaccharides terminating with alpha 2,6- or alpha 2,3-linked sialic acid. The free amino group of the spacer of these oligosaccharides was converted into an isothiocyanate. Its subsequent coupling to bovine serum albumin gave neoglycoproteins with a yield of 2.4-3.6 glycan chains per carrier molecule. This versatile synthetic pathway allows employment of a wide variety of complex-type glycans, which can be introduced to various test systems in vitro and in vivo to evaluate potential biomedical applications. Solid-phase assays with biotinylated sugar receptors revealed discriminatory binding properties of the three neoglycoproteins, especially for the mistletoe lectin. This direct assay system is preferable to the measurement of inhibitory capacities with respect to model ligands. Ligand type- and cell type-dependent quantitative differences in the binding properties of the probes were detected by FACScan analyses with a panel of tumor cell lines and by monitoring of staining in tissue sections for small cell and non-small-cell lung cancer and mesotheliomas. Biodistribution of iodinated neoglycoproteins in mice gave a prolonged presence of the sialylated probes in serum. Relative to the nonasaccharide, the uptake, especially of the iodinated neoglycoprotein with alpha 2,3-sialylated ligand chains, was clearly elevated in mice for kidneys and Ehrlich tumors. On the basis of the documented feasibility of these applications, it is concluded that the further elaboration of glycan chain variants by the described synthetic approach in combination with the given test panel is warranted to evaluate the potential of complex glycan chain-carrying neoglycoproteins for diagnostic and therapeutic purposes.

show abstract

Section: Resultsmentioning

confidence: 99%

Neoglycoproteins with the Synthetic Complex Biantennary Nonasaccharide or Its α2,3/α2,6-Sialylated Derivatives: Their Preparation, Assessment of Their Ligand Properties for Purified Lectins, for Tumor Cells in Vitro, and in Tissue Sections, and Their Biodistribution in Tumor-Bearing Mice

et al. 1997

Self Cite

View full text Add to dashboard Cite

show abstract

“…Preparation of Neoganglioprotein and Lectin-specific AntibodiesThe lysoganglioside of purified GM 1 was obtained by deacylation in 1 M KOH followed by selective re-N-acetylation and covalently linked as sphingosine N-alkyl(sulfosuccinimidyl)ester derivative to carbohydrate-free bovine serum albumin with an incorporation yield of 2-3 units/carrier molecule as described (38,39). Polyclonal antibodies against the mammalian galectins-1 and -3 were raised in rabbits, purified from serum by protein A-Sepharose 4B chromatography, and checked for specificity and lack of cross-reactivity as described elsewhere (40,41).…”

Section: Methodsmentioning

confidence: 99%

Galectin-1 Is a Major Receptor for Ganglioside GM1, a Product of the Growth-controlling Activity of a Cell Surface Ganglioside Sialidase, on Human Neuroblastoma Cells in Culture

Kopitz

Reitzenstein

Burchert

et al. 1998

Journal of Biological Chemistry

262

196

View full text Add to dashboard Cite

Cell density-dependent inhibition of growth and neural differentiation in the human neuroblastoma cell line SK-N-MC are associated with a ganglioside sialidasemediated increase of GM 1 and lactosylceramide at the cell surface. Because these glycolipids expose galactose residues, we have initiated the study of the potential role of galectins in such cellular events. Using specific antibodies, galectin-1 but not galectin-3 was found to be present at the cell surface. Assessment of carbohydratedependent binding revealed a saturable amount of ligand sites approaching 2.6 ؋ 10 6 galectin-1 molecules bound/cell. Presence during cell culture of the sialidase inhibitor 2-deoxy-2,3-dehydro-N-acetylneuraminic acid or of the GM 1 -binding cholera toxin B subunit effected a decrease of the presentation of galectin-1 ligands by 30 -50%. The assumption that GM 1 is a major ligand for galectin-1 was reinforced by the correlation between the number of carbohydrate-dependent 125 I-iodinated GM 1 -neoganglioprotein binding sites and the amount of immunoreactive surface galectin-1, the marked sensitivity of probe binding to the presence of anti-galectin-1 antibody, and the inhibition of cell adhesion to surfaceimmobilized GM 1 by the antibody. The results open the possibility that the carbohydrate-dependent interaction between ganglioside GM 1 and galectin-1 may relay sialidase-dependent alterations in this cell system.Gangliosides can exert a variety of cellular functions that include the triggering and modulation of transmembrane signaling and the mediation of recognition of receptor molecules in homotypic and heterotypic associations (1-4). Owing to this versatility in regulatory processes, it is fitting that the presentation of gangliosides at the cell surface is subject to control mechanisms that involve biosynthesis, endocytic uptake, recirculation, and lysosomal degradation (5, 6). Moreover, the structure of the oligosaccharide chains of gangliosides can be remodeled in situ by the action of a cell surface sialidase in the course of transformation, differentiation and cell contact formation (7-12). In human neuroblastoma cells (SK-N-MC), the activity of this sialidase was directed toward gangliosides with terminal sialic acids, yielding a shift from higher sialylated species to GM 1 and a conversion of GM 3 to lactosylceramide (13). Such alterations of the ganglioside profile are apparently of profound impact on the behavior of the neuroblastoma cells, because the selective inhibition of the ganglioside sialidase activity by 2-deoxy-2,3-dehydro-N-acetylneuraminic acid (NeuAc2en) 1 led to marked changes in cellular morphology, a complete release from contact inhibition of growth, and a loss of differentiation markers (14,15). Because the underlying molecular events are unknown, we have now addressed questions on the presence and nature of potential receptors for GM 1 and/or lactosylceramide that are generated by the action of the ganglioside sialidase on the surface of neuroblastoma cells.The current literature prompts inves...

show abstract

“…logical testing [269][270][271][272]. Coupling of purified glycans from natural sources is thus no longer the only approach to recruit complex oligosaccharides as tools for receptor search [273][274][275][276], whose concepts and successes will be presented in the section on glycan recognition. When no further processing will reduce the size of the Man 5 GlcNAc 2 chain, it is committed to the generation of high-mannose-type glycans, which are common products of yeasts but are also present in animal glycoproteins such as ovalbumin.…”

Section: Review Articlementioning

confidence: 99%

Eukaryotic glycosylation: whim of nature or multipurpose tool?

Reuter¹,

Gabius²

1999

Cellular and Molecular Life Sciences (CMLS)

214

141

View full text Add to dashboard Cite

Protein and lipid glycosylation is a ubiquitous phenomenon. The task of cataloguing the great structural variety of the glycan part has demanded considerable efforts over decades. This patient endeavor was imperative to discern the inherent rules of glycosylation which cannot affirm assumptions on a purely coincidental nature of this type of protein and lipid modification. These results together with theoretical considerations uncover a salient property of oligosaccharides. In comparison with amino acids and nucleotides, monosaccharides excel in their potential to serve as units of hardware for storing biological information. Thus, the view that glycan chains exclusively affect physiochemical properties of the conjugates is indubitably flawed. This original concept has been decisively jolted by the discovery of endogenous receptors (lectins) for distinct glycan epitopes which are as characteristic as a fingerprint or a signature for a certain protein (class) or cell type. Recent evidence documents that these binding proteins are even endowed with the capacity to select distinct low-energy conformers of the often rather flexible oligosaccharides, granting entry to a new level of regulation of ligand affinity by shifting conformer equilibria. The assessment of the details of this recognition by X-ray crystallography, nuclear magnetic resonance spectroscopy, microcalorimetry and custom-made derivatives is supposed to justify a guarded optimism in satisfying the need for innovative drug design in antiadhesion therapy, for example against viral or bacterial infections and unwanted inflammation. This review presents a survey of the structural aspects of glycosylation and of evidence to poignantly endorse the notion that carrier-attached glycan chains can partake in biological information transfer at the level of cell compartments, cells and organs.

show abstract

Carrier-immobilized derivatized lysoganglioside GM1 is a ligand for specific binding sites in various human tumor cell types and peripheral blood lymphocytes and monocytes

Cited by 25 publications

References 22 publications

Neoglycoproteins with the Synthetic Complex Biantennary Nonasaccharide or Its α2,3/α2,6-Sialylated Derivatives: Their Preparation, Assessment of Their Ligand Properties for Purified Lectins, for Tumor Cells in Vitro, and in Tissue Sections, and Their Biodistribution in Tumor-Bearing Mice

Neoglycoproteins with the Synthetic Complex Biantennary Nonasaccharide or Its α2,3/α2,6-Sialylated Derivatives: Their Preparation, Assessment of Their Ligand Properties for Purified Lectins, for Tumor Cells in Vitro, and in Tissue Sections, and Their Biodistribution in Tumor-Bearing Mice

Galectin-1 Is a Major Receptor for Ganglioside GM1, a Product of the Growth-controlling Activity of a Cell Surface Ganglioside Sialidase, on Human Neuroblastoma Cells in Culture

Eukaryotic glycosylation: whim of nature or multipurpose tool?

Contact Info

Product

Resources

About