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1.
An antiserum raised against -fructosidase isolated from the cell walls of suspension-cultured carrot cells cross-reacts with many plant proteins and hemocyanin ofHelix pomatia. The shared epitope appears to be a small complex glycan with a (1–2)-linked xylose residue attached to the -linked mannose residue of the core of an asparagine-linked oligosaccharide. There is strong cross-reactivity with the proteins of many seed plants, molluscs and insects, and no cross-reactivity with the proteins of fungi, algae, mosses, ferns, or any of the vertebrates tested. Xylose-containing glycans appear to increase the immunogenicity of the proteins to which they are attached, and we suggest that they may be responsible for some allergic responses of people that are repeatedly exposed to plant or insect proteins. 相似文献
2.
Henri Debray Danuta Dus Jean-Michel Wieruszeski Gérard Strecker Jean Montreuil 《Glycoconjugate journal》1991,8(1):29-37
Four bi-antennary glycan fractions of theN-acetyllactosamine-type, derived from a Lewis lung carcinoma (LL2) cell subline resistant to theAleuria aurantia agglutinin were studied by 400 MHz1H-NMR spectroscopy. By this method, their antennae were found to be terminated either by (2-3 or 6)-linkedN-acetylneuraminic acid or (1-3)-linked galactose residues. The primary structure of glycans of these four glycopeptide or derived oligosaccharide-alditols has been determined in full detail.Abbreviations NAc
N-acetyl group
- NGc
N-glycolyl group
- GlcNAc
N-acetylglucosamine
- NeuAc
N-acetylneuraminic acid
- NeuGc
N-glycolylneuraminic acid
- Man
mannose
- Gal
galactose
- Fuc
fucose
- Con A
concanavalin A
- LCA
Lens culinaris agglutinin
- AAA
Aleuria aurantia agglutinin
- WGA
Wheat germ agglutinin
- RCA II
Ricinus communis agglutinin II
- PBS
phosphate buffered saline, 0.01m Na2HPO4/0.14m NaCl, pH 7.2
- HPLC
high performance liquid chromatography
- EMEM
Eagle's Minimal Essential Medium
- LecR
lectin resistant
- MG
-methylglycoside 相似文献
3.
Keiko Tadano-Aritomi Harumi Kubo Philip Ireland Takeshi Kasama Shizuo Handa Ineo Ishizuka 《Glycoconjugate journal》1996,13(2):285-293
A novel mono-sulfated glycosphingolipid based on the gangliotriaose core structure was isolated from rat kidney. The isolation procedure involved extraction of lipids with chloroform/methanol, mild alkaline methanolysis, column chromatographies with anion exchangers and silica beads. The structure was characterized by compositional analysis, FTIR spectroscopy, methylation analysis,1H-NMR spectroscopy and negative-ion liquid secondary ion mass spectrometry (LSIMS) using the intact glycolipid and its desulfation product. The two dimensional chemical shift correlated spectroscopy provided information on the sugar sequence as well as anomeric configurations, and indicated the presence of a 3-O-sulfatedN-acetylgalactosamine within the molecule. Negative-ion LSIMS with high- and low-energy collision-induced dissociation defined the sugar sequence and ceramide composition, confirming the presence of a sulfatedN-acetylgalactosamine at the non-reducing terminus. From these results, the complete structure was proposed to be HSO3-3GalNAc1-4Gal1-4Glc1-1Cer (Gg3Cer III3-sulfate, SM2b).
Abbreviations: Abbreviations for sulfated glycolipids [17] follow the modifications of the nomenclature system of Svennerholm for gangliosides [37], and the designation of the other glycosphingolipids follows the IUPAC-IUB recommendations [38]. Cer, ceramide; LacCer, lactosylceramide, Gal1-4Glc1-1Cer; Gg3Cer, gangliotriaosylceramide, GalNAc1-4Gal1-4Glc1-1Cer; Gg4Cer, gangliotetraosylceramide, Gal1-3GalNAc1-4Gal1-4Glc1-1Cer; iGb4Cer, isoglobotetraosylceramide, GalNAc1-3Gal1-3Gal1-4Glc1-1Cer; Gb4Cer, globotetraosylceramide, GalNAc1-3Gal1-4Gal1-4Glc1-1Cer; SM4s, galactosylceramide sulfate, GalCer I3-sulfate; SM3, lactosylceramide sulfate, LacCer II3-sulfate; SM2a, Gg3Cer II3-sulfate; SM2b, Gg3Cer III3-sulfate; SB2, Gg3Cer II3,III3-bis-sulfate; SM1a, Gg4Cer II3-sulfate; SM1b, Gg4Cer IV3-sulfate; SB1a, Gg4Cer II3,IV3-bissulfate; GLC, gas-liquid chromatography; GC-MS, gas chromatography-mass spectrometry; DQF, double quantum filtered; COSY, chemical-shift-correlated spectroscopy; LSIMS, liquid secondary ion mass spectrometry; CID, collision-induced dissociation; MS/MS, tandem mass spectrometry. 相似文献
4.
Structural characterization of sulfated and sialyl Lewis (Le)-type glycosphingolipids performed by fast atom bombardment (FAB) and electrospray ionization (ESI) mass spectrometry is described. Both FAB and ESI collision-induced dissociation tandem mass spectrometry (CID-MS/MS) of acidic glycosphingolipids allowed identification of the sulfated or sialyl sugar, and provided information on the saccharide chain sequence. The negative-ion tandem FABMS of sulfated Le-type glycosphingolipids having the non-reducing end trisaccharide ion as the precursor can be used to differentiate the Lea- and LeX-type oligosaccharides. The ESI CID-MS/MS of multiple-charged ions provided even more detailed structural information, and some of the useful daughter ions appeared with higherm/z values than the precusor because of a lower charge-state. These methodologies can be applied to the structural analyses of glycoconjugates with much larger molecular masses and higher polarity, such as the poly-sulfated and sialyl analogues.Abbreviations CID
collision-induced dissociation
- ESI
electrospray ionization
- FABMS
fast atom bombardment mass spectrometry
- Fuc
fucose
- Gal
galactose
- GlcNAc
N-acetylglucosamine
- Le
Lewis
- Lea
Lewisa
- LeX
LewisX
- MS/MS
mass spectrometry/mass spectrometry
- NeuAc
N-acetylneuraminic acid
- 3-SO4-Lea
3-sulfated Lea pentaosyl ceramide
- 3-SO4-LeX
3-sulfated LeX pentaosyl ceramide
- 2,3-SO4-LeX
2,3-disulfated LeX pentaosyl ceramide
- 3-S-Lea
3-sialyl Lea pentaosyl ceramide
- 3-S-Lex
3-sialyl LeX heptaosyl ceramide
- 3-S-LeX-LeX
3-sialyl-Lex-Lex octaosyl ceramide. 相似文献
5.
Ana Maria Landeira-Fernandez Maricilia S. Costa Leopolde de Meis 《Bioscience reports》1996,16(6):439-451
Vesicles derived from maize roots retain a membrane bound H+-ATPase that is able to pump H+ at the expense of ATP hydrolysis. In this work it is shown that heparin, fucose-branched chondroitin sulfate and dextran sulfate 8000 promote a shift of the H+-ATPase optimum pH from 6.0 to 7.0. This shift is a result of a dual effect of the sulfated polysaccharides, inhibition at pH 6.0 and activation at pH 7.O. At pH 6.0 dextran 8000 promotes an increase of the apparent Km for ATP from 0.28 to 0.95 mM and a decrease of the Vmax from 14.5 to 7.1 mol Pi/mg · 30 min–1. At pH 7.0 dextran 8000 promotes an increase in Vmax from 6.7 to 11.7 mol Pi/mg · 30 min–1. In the presence of lysophosphatidylcholine the inhibitory effect of the sulfated polysaccharides observed at pH 6.0 was not altered but the activation of pH 7.0 decreased. It was found that in the presence of sulfated polysaccharides the ATPase became highly sensitive to K+ and Na+. Both the inhibition at pH 6.0 and the activation promoted by the polysaccharide were antagonized by monovalent cations (K+>Na+Li+).Abbreviations Mops
4-morpholinopropanesulfonic acid
- EDTA
ethylenediaminetetraacetic acid
- ACMA
9-amino-6-chloro-2-methoxyacridine
- FCCP
carbonyl cyanide p(trifluoromethoxy)-phenylhyrazone 相似文献
6.
Complete acid hydrolysis of red algal galactans in the presence of borane - 4-methylmorpholine complex has been shown to prevent the acid degradation of 3,6-anhydrogalactose derivatives by their reduction to the corresponding 3,6-anhydro-galactitols, whereas all the other monosaccharides are liberated essentially in the non-reduced form; the reductive hydrolysis products may be determined quantitatively using gas-liquid chromatography (GLC). The method is recommended for preliminary characterization of the polysaccharide composition of red algal biomass. Partial acid hydrolysis of galactans in the presence of the same reducing agent gives rise to reduced oligosaccharides having terminal 3,6-anhydrogalactitol residues. Based on this reaction, the attribution of unknown galactans to the agar or carrageenan groups is possible by partial reductive hydrolysis of small samples of algal biomass with subsequent identification of agarobiitol or carrabiitol acetates by GLC. Sulfate groups are substantially retained under partial reductive hydrolysis conditions; the isolation by liquid chromatography and elucidation of structures of reduced sulfated oligosaccharides may be of great value for the structural analysis of complex red algal galactans. 相似文献
7.
Three major glycan fractions of 580 kDa (g580), 150 kDa (g150), and 2 kDa (g2) were isolated and purified from Lytechinus pictus sea urchin embryos at the mesenchyme blastula stage by gel filtration and high pressure liquid chromatography. Chemical analysis, by gas chromatography, revealed that g580 is highly sulfated and rich in N-acetylglucosamine, N-acetylgalactosamine, glucuronic acid, and fucose. The g150 fraction is less acidic than g580 and contains high amounts of amino sugars, xylose, and mannose. The g2 fraction is neutral, rich in N-acetylglucosamine, mannose, and galactose. The g580 and g150 fractions are resistant to glycosaminoglycan-degrading enzymes, indicating that they are distinct from the glycosaminoglycans. The g580 fraction resembles, with respect to chemical composition, a previously characterized 200 kDa sponge adhesion glycan (g200). The binding of the monoclonal antibody Block 2, which recognizes a repetitive epitope on g200, as well as of the anti-g580 polyclonal antibodies to both g580 and g200 indicated that these two glycans share similar antigenic determinants. The Fab fragments of the Block 2 antibody, which previously have been shown to inhibit cell adhesion in sponges, also blocked the reaggregation of dissociated sea urchin mesenchyme blastula cells. These results indicate that g580 carries a carbohydrate epitope, similar to the sponge adhesion epitope of g200, which is involved in sea urchin embryonal cell adhesion. 相似文献
8.
Loïc Faye Kenneth D. Johnson Arnd Sturm Maarten J. Chrispeels 《Physiologia plantarum》1989,75(2):309-314
In plants, glycoproteins with asparagine-linked glycans (oligosaccharides) are found in vacuoles, in the extracellular space or matrix, and associated with the endo-membrane system (endoplasmic reticulum, Golgi apparatus, plasma membrane, tonoplast). These glycans are of the high-mannose type, with a structure identical to that found in other organisms (mammals, yeast), or of the complex type with a β1–2 linked xylosyl residue not found in mammalian complex glycans. Asparagine-linked glycans play multiple roles by modifying the physicochemical properties of the polypeptides to which they are attached. 相似文献
9.
Peter Jackson 《Molecular biotechnology》1996,5(2):101-123
The glycans of glycoconjugates mediate numerous important biological processes. Their separation and structural determination
present considerable difficulties because of the small quantities that are available from biological sources and the inherent
difficulty of analyzing the wide variety of complex structures that exist. A method for the analysis of reducing saccharides
by PAGE that uses specific fluorophore labeling and is simple, rapid, sensitive, and readily available to biological researchers,
has been developed. The method is known acronimically either as PAGEFS (PAGE of Fluorophore-labeled Saccharides) or in one
commercial format as FACE (Fluorophore-Assisted Carbohydrate Electrophoresis). In the PAGEFS method, saccharides having an
aldehydic reducing end group are labeled quantitatively with a fluorophore and then separated with high resolution by PAGE.
Two fluorophores, 8-aminonaphthalene-l,3,6-trisulfonic acid (ANTS) and 2-aminoacridone (AMAC), have been used to enable the
separation of a variety of saccharide positional isomers, anomers, and epimers. Subpicomolar quantities of individual saccharides
can be detected using a sensitive imaging system. Mixtures of oligosaccharides obtained by enzymatic cleavage from glycoproteins
can be labeled and electrophoresed to yield an oligosaccharide profile of each protein. AMAC can be used to distinguish unequivocally
between acidic and neutral oligosaccharides. Methods for obtaining saccharide sequence information from purified oligosaccharides
have been developed using enzymatic degradation. Other applications and the potential of the system are described. 相似文献
10.