The gas chromatographic properties of biologically important N-acetylglucosamine derivatives, monosaccharides, disaccharides, trisaccharides, tetrasaccharides and pentasaccharides

The diffusion of fluorescein isothiocyanate-labclled dextran molecules in suspensions of centrifugally, tightly packed, erythrocyte ghosts was measured by fluorescence recovery after photobleaching. In comparison with diffusion in aqueous solution, the diffusion coefficients for probe molecules of varying size were about two orders of magnitude smaller. It was established that the dextran molecules remained in the space between the ghosts. Since crosslinking membrane surface carbohydrates with antibodies further inhibits diffusion, it is assumed that interactions between surface carbohydrates and the probe molecules are the cause of slow diffusion. Two alternative models are discussed.Intercellular diffusion; FRAP; FITC-dextran; Cell membrane carbohydrate; Carbohydrate interaction; (Erythrocyte ghost)

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“…Protein [9], sialic acid [10] and the content of various monosaecharides [11] were estimated using standard procedures.…”

Section: Analytical Proceduresmentioning

confidence: 99%

Diffusion of fluorescein‐labelled molecules in suspensions of erythrocyte ghosts

1988

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“…For glycoconjugate analysis, the biopsy specimens were homogenized in water and analyzed for monosaccharides by gas-liquid chromatography as described previously (17)(18)(19)) and for protein as described by Hartree (20). A Perkin-Elmer Model 900 gas chromatograph, with hydrogen flame ionization detectors and nitrogen as carried gas, was used.…”

Section: Patientsmentioning

confidence: 99%

Atrophic gastritis in sjögren's syndrome. Morphologic, biochemical, and immunologic findings

Maury

Törnroth

Teppo

1985

Arthritis & Rheumatism

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Gastric studies were carried out in 16 patients with well‐documented Sjögren's syndrome (SS), 43 matched rheumatic disease patients without SS, and 7 patients with chronic atrophic gastritis not associated with SS. Chronic atrophic gastritis was a much more common finding in the SS patients than in the rheumatic disease control patients. Significant hypopepsinogenemia was present in 11 of 16 SS patients. In 6 patients this was combined with hypergastrinemia, a combination highly specific for chronic atrophic gastritis. The lowest pepsinogen levels were seen in patients with primary SS associated with high levels of SS‐B antibody. On a histologic and biochemical basis, it was not possible to distinguish the gastric findings in primary SS from those in secondary SS, nor to distinguish chronic atrophic gastritis associated with SS from that not associated with SS. We conclude that chronic atrophic gastritis is a prominent feature in SS and that the severity of the gastritis appears to correlate with some serologic parameters of SS.

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“…Quantification of glycolipids isolated from labeled erythrocytes was done by gas-liquid chromatography after methanolysis as described by Bhatti et al [13]. For the determination of the degree of deuteration of the N-acetylgalactosamine, the trimethylsilyl derivatives were analyzed by gasliquid chromatography/mass spectrometry (Hewlett-Packard 3992A) on a column of 4% OV-101 with a starting temperature of 180°C.…”

Section: Gas-liquid Chrornatography/mass Spectrometry Of Sugarsmentioning

confidence: 99%

Exposure of the major human red-cell glycolipid, globoside, to galactose oxidase

Lampio¹,

Finne

Homer

et al. 1984

Eur J Biochem

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Plasma membrane glycolipids are localized at the outer leaflet of the lipid bilayer, and their carbohydrate portions are exposed to the environment. The efficiency of exposure has, however, not been known. We have been able to determine the availability of the major red cell glycolipid, gIoboside, to externally added galactose oxidase. Red cells were extensively treated with the enzyme and the oxidized cells reduced with NaBD4. After isolation the extent of exposed globoside was estimated by mass spectrometry. The results show that the exposure of globoside varies in red cells of different individuals from 37 -66%. The fatty acid composition of externally available globoside was the same as that of non-oxidized globoside. The exposure was not influenced by protease treatment of intact cells and no correlation was found with different ABO blood groups.Glycolipids are ubiquitous mammalian cell-surface components. Their carbohydrate head-groups can act as receptors for antibodies, lectins, bacterial toxins, and microbes [l]. They may also pIay a role in cell-cell interactions.Several studies have demonstrated that glycolipids in plasma membranes are localized at the outer cell surface and no glycolipids are found at the cytoplasmic aspect [2, 31. There is, however, evidence that glycolipids are only partially exposed to the environment [4]. The crypticity may vary during the cell cycle and by transformation [5]. Essentially nothing is, however, known about the efficiency of exposure [l].It would obviously be important to determine the degree of exposure and its variation under different conditions. We have therefore treated human erythrocytes with galactose oxidase until all accessible glycolipid was oxidized, and thereafter reduced the cells with NaBD4. The glycolipids were isolated and the ratio of surface lipid to inaccessible lipid determined by mass spectrometry. By using this technique we show that only part of the major glycolipid, globoside, is exposed in human erythrocyte membranes, and that the degree of exposure differs in different individuals. MATERIALS AND METHODS CellsBlood samples of known ABO blood groups were taken into heparinized tubes, and the erythrocytes were washed by centrifugation four times with 0.15 M NaCl-0.01 M sodium phosphate, pH 7.4 (NaC1/PO4). All labeling experiments were done using freshly drawn cells. EnzymesIn the early experiments galactose oxidase (Dactylium dendroides) was obtained from Kabi Ltd (Stockholm, Enzymes. Galactose oxidase (EC 1 .I -3.9); horseradish peroxidase (EC 1.11.1.7); catalase (EC 1.11.1.6); trypsin (EC 3.4.21.4); Streptomyces griseus protease (EC 3.4.24.4); Vibrio cholerae neuraminidase (EC 3.2.1.18). Sweden), later galactose oxidase (type V) from Sigma Chemical Co. (St. Louis, MO) was used. The enzyme was heated for 30 min at 56°C before use to destroy possibly contaminating proteases [3]. No difference was found between the preparations. Horseradish peroxidase (type II), bovine liver catalase, trypsin from bovine pancreas (type XI) and Streptomyces gris...

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The gas chromatographic properties of biologically important N-acetylglucosamine derivatives, monosaccharides, disaccharides, trisaccharides, tetrasaccharides and pentasaccharides

Cited by 291 publications

References 14 publications

Diffusion of fluorescein‐labelled molecules in suspensions of erythrocyte ghosts

Diffusion of fluorescein‐labelled molecules in suspensions of erythrocyte ghosts

Atrophic gastritis in sjögren's syndrome. Morphologic, biochemical, and immunologic findings

Exposure of the major human red-cell glycolipid, globoside, to galactose oxidase

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