Papers by Herbert Wiegandt
Hoppe-Seyler´s Zeitschrift für physiologische Chemie, 1974
Lipids, 1994
ABSTRACT A method is described for the preparation of ganglioside Glac2 [(II3(NeuAc)2-LacCer, GD3... more ABSTRACT A method is described for the preparation of ganglioside Glac2 [(II3(NeuAc)2-LacCer, GD3] from cream of bovine milk using liquid-phase extraction with methanol or ethanol followed by anion exchange chromatography. The method is rapid and inexpensive; 1 kg cream, centrifuged from 14-15 L of bovine milk, yields approximately 70 mg of pure ganglioside Glac2. The sialic acid constituent of ganglioside Glac2 isolated from bovine milk cream consists solely of the N-acetylneuraminic acid derivative. The major components of its ceramide consist of octadecasphing-4-enine and the 22:0 (behenic acid) and 23:0 fatty acids.
Hoppe-Seyler´s Zeitschrift für physiologische Chemie, 1982
We studied the mode of uptake of exogenous sialoglycolipids with a permanent mouse cell line (Clo... more We studied the mode of uptake of exogenous sialoglycolipids with a permanent mouse cell line (Clone-1D) and compared the cell association of ganglioside II3 NeuAc-Gg4 Cer with that of a synthetic one-C18-hydrocarbon-chain ganglioside analogue (desphingo-G1-stearamide). In contrast to monolayer cultures, Clone-1D cells in suspension bind both sialo-glycolipids rapidly and independently of time and temperature (4 degrees/37 degrees C) of incubation. Three modes of cell association were distinguished: 1) a serum-and serum albumin-sensitive association; 2) a trypsin-sensitive association and 3) a serum- and trypsin-insensitive association. Inhibition of pinocytosis by deoxyglucose/sodium azide did not significantly influence cellular uptake or release of exogenous ganglioside. In monolayer culture, Clone 1D cells in M-phase associate more ganglioside than randomly growing cells. This may explain in part the inhomogeneous cellular distribution of ganglioside binding. In contrast, uptake of desphingo-G1 -stearamide by cells in mitosis equalled that bound by control cells.
Molecular and Biochemical Parasitology, 1992
ABSTRACT A comparative survey was undertaken of the neutral fraction glycolipids from the metaces... more ABSTRACT A comparative survey was undertaken of the neutral fraction glycolipids from the metacestodes of 3 taeniid species, Taenia crassiceps, Taenia solium and Taenia saginata, to determine their chemical and serological staining patterns on separation by thin-layer chromatography. The orcinol-positive patterns of T. solium and T. saginata metacestodes exhibited a closer superficial resemblance to each other than to T. crassiceps or T. saginata adults. A comparison of component migration properties against standards of known structure indicated the main oligosaccharide chains to be mono-, di-, tri- and tetrasaccharides; however, in T. solium this was extended to at least a heptasaccharide. The multiple banding characteristic of each component is a consequence of lipid moiety heterogeneity. Serologically, the patterns of the 3 taeniid species neutral fraction glycolipids showed virtually the same immunological reactivity towards mouse normal serum, infection serum and a monospecific, polyclonal antibody directed against the trisaccharide component of T. crassiceps. The latter antibody was isolated from mouse infection serum by affinity chromatography on a column of glycolipid-bound octyl-Sepharose CL-4B. Immunochemically, the major common epitope expressed by the neutral fraction glycolipids of the 3 taeniid species is the same or very similar to the glycosphingolipid, neogalatriaosyl ceramide derived from the marine mollusc Turbo cornutus (Gal(beta 1-6) Gal(beta 1-6) Gal(beta 1-1)Cer). Host tissue neutral fraction glycolipids, porcine muscle and bovine muscle, as well as human spleen, were not immunoreactive.
Hoppe-Seyler's Zeitschrift für physiologische Chemie, 1978
The stimulation by cholera toxin of adenylate cyclase in Chinese hamster ovarian cells could be i... more The stimulation by cholera toxin of adenylate cyclase in Chinese hamster ovarian cells could be inhibited by various ligands. The latter have been shown to contain the structural oligosaccharide entities required for binding to cholera toxin, established as Galbeta1 leads to 3GalNAcbeta1 leads to 4Gal3 comes from 2alphaNeuAc. The different inhibitory potency of the ligands thereby correlates with the size of the aggregates formed with the toxin, which in turn depends on the valency of the ligands. The conclusion is drawn from a comparison of the interaction of cholera toxin and its B-protomer with ganglioside II3NeuAc-GgOse4-Cer, the newly synthesized bis-(monosialo-gangliotetraityl)amine and monosialogangliotetraose. In a double diffusion test cholera toxin B-protomer precipitated with the ganglioside II3 NeuAcGgOSE4-Cer and the divalent ligand bis(monosialo-gangliotetraityl)amine, suggesting the formation of high molecular weight aggregates, whereas no precipitation was observed w...
The Journal of biological chemistry, Jan 2, 2007
Ceramides are vital components of the water barrier in mammalian skin. Epidermis-specific, a majo... more Ceramides are vital components of the water barrier in mammalian skin. Epidermis-specific, a major ceramide portion contains omega-hydroxy very long chain fatty acids (C30-C36). These omega-hydroxy ceramides (Cers) are found in the extracellular lamellae of the stratum corneum either as linoleic acyl esters or protein bound. Glucosylceramide is the major glycosphingolipid of the epidermis. Synthesized from ceramide and UDP-glucose, it is thought to be itself an intracellular precursor and carrier for extracellular omega-hydroxy ceramides. To investigate whether GlcCer is an obligatory intermediate in ceramide metabolism to maintain epidermal barrier function, a mouse with an epidermis-specific glucosylceramide synthase (Ugcg) deficiency has been generated. Four days after birth animals devoid of GlcCer synthesis in keratinocytes showed a pronounced desquamation of the stratum corneum and extreme transepidermal water loss leading to death. The stratum corneum appeared as a thick unst...
Hybridoma, 1993
A cloned mouse hybridoma was established that secreted a monoclonal antibody directed against cer... more A cloned mouse hybridoma was established that secreted a monoclonal antibody directed against certain neutral glycosphingolipids of the third-instar larvae of Calliphora vicina (Insecta: Diptera). The isotype of the designated monoclonal antibody CNF-I (Calliphora Neutral Fraction) was determined as IgG3. By the use of purified neutral glycosphingolipids of C. vicina, qualitative thin-layer chromatography immunostaining and semi-quantitative enzyme-linked immunosorbent assay determinations, the epitope was specified as a terminal alpha 1-4-linked N-acetylgalactosamine to subterminal N-acetylgalactosamine, which is present on the components GalNAc alpha 4GalNAc beta 4GlcNAc beta 3Man beta 4Glc beta lCer (N5a) and GalNAc alpha 4GalNAc beta 4-(6' PEtn-)GlcNAc beta 3Man beta 4Glc beta lCer (Nz5a). After enzymatic removal of the terminal alpha-N-acetylgalactosamine residue, the epitope reactivity was destroyed. The distribution of the CNF-I recognized epitope among neutral glycolipid...
Advances in experimental medicine and biology, 1980
ABSTRACT Gangliosides have been found to contain up to five sialic acid moieties bound to galacto... more ABSTRACT Gangliosides have been found to contain up to five sialic acid moieties bound to galactose residues in α, 2->3 linkages either at the end of the oligosaccharide chain or within the chain, and may form di- and trisialyl groups with α,2->8 linkages (1,2). N-Acetylneuraminic acid (NeuAc) and N-glycolylneuraminic acid (NeuG1) have been identified as the commonly occurring sialic acids in gangliosides, while the O-acetylated sialic acids occur less frequently. The 9-O-acetyl-N-acetylneuraminic acid has been found in gangliosides from a variety of vertebrates including man (3), and 4-O-acetyl-N-glycolylneuraminic acid (4-OAc-NeuGl) has been isolated from horse erythrocyte hematoside (4).
Advances in experimental medicine and biology, 1980
European journal of biochemistry / FEBS, 1974
It was confirmed that ganglioside G Gtet 1, ie Galβ1→ 3GalNAcβ1→ 4 [NeuNAcα2→ 3] Galβ1→ 4Glc-cera... more It was confirmed that ganglioside G Gtet 1, ie Galβ1→ 3GalNAcβ1→ 4 [NeuNAcα2→ 3] Galβ1→ 4Glc-ceramide, specifically interacts with choleragen (cholera-cxoenterotoxin) as shown by precipitate formation and inhibition of the toxicity. The isolated carbohydrate ...
Hybridoma, 1990
A cloned, hybridoma cell-line was established that secreted the monoclonal antibody CAF-I followi... more A cloned, hybridoma cell-line was established that secreted the monoclonal antibody CAF-I following stimulation of the donor Balb/c mouse spleen cells by the total acidic fraction glycolipids of the third-instar larvae of Calliphora vicina (Insecta:Diptera). The monoclonal antibody isotype was IgG3 By qualitative (TLC-immunostaining) and semi-quantitative (enzyme-linked immunosorbent assay) methods, and comparison with the cross-reactivity of known monoclonal antibodies, the epitope was specifically located on the terminal, non-reducing end of the oligosaccharide chain of most of the insect acidic glycolipids. Following isolation of the two main acidic glycolipids of C. vicina larvae (A5c and Az5c), exoglycosidase treatment characterized the terminal disaccharide CAF-I epitope as glucuronic acid bound to subterminal galactose, both in the beta-anomeric configuration: G1cA beta-Ga1 beta-. The immunohistological distribution of this epitope in the dipteran, Drosophila melanogaster, sh...
Hoppe-Seyler´s Zeitschrift für physiologische Chemie, 1976
The fixation of cholera toxin by ganglioside GGtet1 is dependent on the nature of the carbohydrat... more The fixation of cholera toxin by ganglioside GGtet1 is dependent on the nature of the carbohydrate as well as the lipid moiety of the glycolipid. The role of the lipid in binding to the toxin investigated with synthetic ganglioside analogues (gangliosidoides). The interaction between glycolipid and toxin was followed by precipitate formation, by inhibition of toxicity and in polyacrylamide gel electrophoresis. For specific precipitation, an aliphatic hydrocarbon chain at least 14 C-atoms in length is required. Some of the gangliosidoides form high molecular weight complexes with cholera toxin at lower molar ratios of ligand to protein than the natural compound. None of the synthetic gangliosidoides equalled natural ganglioside in its ability to inhibit the effects of the toxin in vivo, but some did show considerable inhibitory activity ih monosialo-gangliotetraose or corresponding sialo-glycolipids prevents the easy degradation of the B-protein of cholera toxin into protein subunits by sodium dodecylsulfate.
European Journal of Biochemistry, 1999
From the edible mushroom, the basidiomycetes Agaricus bisporus and Agaricus campestris, a novel c... more From the edible mushroom, the basidiomycetes Agaricus bisporus and Agaricus campestris, a novel carbohydrate-homologous series of four glyco-inositol-phospho-sphingolipids, designated basidiolipids, was isolated and the constituents purified. The chemical structures of the basidiolipids were elucidated to be: Manpbeta1-2inositol1-phospho-ceramide, Galpalpha-6[Fucpalpha-2]Galpbeta-6Manpbeta-2i nositol1-phospho-ceramide, Galpalpha-6Galpalpha-6[Fucpalpha-2]Galpbeta- 6Manpbeta-2inositol1-phospho-ceramide and Galpalpha-6Galpalpha-6Galpalpha-6[Fucpalpha-2] Galpbeta-6Manpbeta-2ino sitol1-phospho-ceramide. All four glycolipids contained a ceramide which was composed of phytosphingosine and predominantly alpha-hydroxy-behenic and alpha-hydroxy-lignoceric acid.
European Journal of Biochemistry, 1991
A group of Calliphora vicina pupal glycolipids could be segregated from the neutral glycosphingol... more A group of Calliphora vicina pupal glycolipids could be segregated from the neutral glycosphingolipids, according to their two-dimensional TLC migration properties and positive reactions toward ninhydrin and fluorescamine spray reagents. These classified zwitterionic glycolipids were isolated by silica-gel column chromatography and characterized by the presence of a N-acetyl-glucosamine-bound phosphoethanolamine residue. The structural elucidation of the oligosaccharide moieties was performed by the determination of constituent carbohydrates as alditol acetates, linkage analysis by permethylation, exoglycosidase cleavage, fast-atom-bombardment mass spectrometry and NMR spectroscopy. The dominant fatty acid and sphingoid base species of the ceramide moieties were C20:0 (arachidic acid) and C14:1 (tetradecasphing-4-enine), respectively. The chemical structures of the zwitterionic, biogenetic glycosphingolipid series were determined as: (PEtn-6')GlcNAc(beta 1-3)Man(beta 1-4)Glc beta Cer; GalNAc(beta 1-4)(PEtn-6')GlcNAc(beta 1-3)Man(beta 1-4)Glc beta Cer; GalNAc(alpha 1-4)GalNAc(beta 1-4)(PEtn-6')GlcNAc(beta 1-3)Man(beta 1- 4)Glc beta Cer; Gal(beta 1-3)GalNAc(beta 1-4)(PEtn-6')GlcNAc(beta 1-3)Man(beta 1-4)Glc beta Cer; Gal(beta 1-3)GalNAc(alpha 1-4)GalNAc(beta 1-4)(PEtn-6')GlcNAc(beta 1- 3)Man(beta 1-4)Glc beta Cer; GlcNAc(beta 1-3)Gal(beta 1-3)GalNAc(alpha 1-4)GalNAc(beta 1-4)(PEtn- 6')GlcNAc(beta 1-3)Man(beta 1-4)Glc beta Cer.
European Journal of Biochemistry, 1992
The presence of glycosphingolipids in the metacestodes of the fox tapeworm, Tueniu crassiceps, ha... more The presence of glycosphingolipids in the metacestodes of the fox tapeworm, Tueniu crassiceps, has been established. The normal-phase TLC pattern of the neutral-fraction glycolipids revealed groups of bands corresponding to homologous components of increasing sugar chain length. The three simplest glycolipid components have been isolated and their chemical constitution determined as being of the neogala series: Galpl Cer, Galp6Galpl Cer and Galp6Galfl6Galbl Cer. The ceramide tetrasaccharide fraction has been found to consist of a mixture of neogalatetraosylceramide, as an elongation of the neogala series, Gal/36Galp6Galp6GalplCer and the component Gala4Galp6-GalB6GalplCer (both occurring in approximately equimolar proportions). The long-chain bases of the ceramide monogalactoside, digalactoside, trigalactoside and tetragalactosides contain, as well as small amounts of sphingosine, predominantly dihydrosphingosine/phytosphingosine in the approximate ratios 1.7: 1, 1.4: 1 , l : 1 and 2.3: 1, respectively. The major ceramide fatty acids have particularly long chains, with hexacosanoic and octacosanoic acids predominating. Upon reverse-phase TLC, the glycolipid components ceramide monogalactoside, digalactoside and trigalactoside were each separable into five component bands. Parent glycolipid components therefore show component band distributions comparable to one another in being governed by similar ceramide constitutions.
European Journal of Biochemistry, 2001
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Papers by Herbert Wiegandt