Separation on Dowex 50 Ion-Exchange Resin of Glucosamine and Galactosamine and their Quantitative Determination.

Gardell, S.; Storgårds, Torsten; Zachariasen, Hans; Westermark, Henrik; Steensgaard, Inger; Rosenberg, Th.

doi:10.3891/acta.chem.scand.07-0207

Cited by 666 publications

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“…Anhydromannose was determined by the indole method described by Dische & Borenfreund (1950). Hexosamine analyses were performed by a modification (Gardell, 1953) of the Elson-Morgan reaction, after hydrolysis of saccharides in 4M-HCl at 100°C for 14h. Glucosamine/galactosamine ratios were measured by chromatography on a column of Aminex A-5 (Bio-Rad Laboratories, Richmond, Calif., U.S.A.), as described by Lohmander (1972).…”

Section: Methodsmentioning

confidence: 99%

Distribution of sulphate and iduronic acid residues in heparin and heparan sulphate

Höök

Lindahl

Iverius

1974

Biochemical Journal

120

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1. A method was developed for determination of the uronic acid composition of heparin-like glycosaminoglycans. Polymers or oligosaccharides are degraded to monosaccharides by a combination of acid hydrolysis and deamination with HNO2. The resulting uronic acid monosaccharides (accounting for about 70% of the uronic acid contents of the starting materials) are isolated and converted into the corresponding aldono-1,4-lactones, which are separated by g.l.c. The calculated ratios of glucuronic acid/iduronic acid are reproducible within 5 %. 2. Samples of heparin from pig intestinal mucosa (molar ratio of sulphate/disaccharide unit, 2.40) and heparan sulphate from human aorta (sulphate/disaccharide ratio, 0.46) were subjected to uronic acid analysis. L-Iduronic acid constituted 77 % and 19 % respectively of the total uronic acid contents. 3. The correlation between the contents of sulphate and iduronic acid indicated by this finding also applied to the fractionated deamination products of the two polymers. The sulphated fragments varied in size from disaccharide to octasaccharide (or larger) and showed sulphate/disaccharide molar ratios in the range of 0.05-2.0. The proportion of iduronic acid increased with increasing ester sulphate contents of the oligosaccharides. 4. Previous studies on the biosynthesis of heparin in a cell-free system have shown that L-iduronic acid residues are formed by C-5 epimerization ofD-glucuronic acid units at the polymer level; the process requires concomitant sulphation of the polymer. The results obtained in the present structural study conform to these findings, and suggest further that similar mechanisms may operate in the biosynthesis of heparan sulphate. The epimerization reaction appears to be linked to the sulphation of hydroxyl groups but does not seem to require sulphation of the target uronic acid residues. The significance of sulphamino groups in relation to the formation of iduronic acid is unknown.

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Section: Methodsmentioning

confidence: 99%

Distribution of sulphate and iduronic acid residues in heparin and heparan sulphate

Höök

Lindahl

Iverius

1974

Biochemical Journal

120

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“…The salt-free solutions were evaporated to dryness in a flash evaporator and the residues were taken up in water and centrifuged. The concentrations of uronie acid (8), hexosamines (9), and nitrogen (10) in these superuatant solutions were determined. In some of the solutions, after oxidation with nitric acid, the concentration of sulfur was determined by a modification of the conductometrie procedure of Paulson (11).…”

Section: Turnover Of S~-sul~'at~mentioning

confidence: 99%

Turnover of S35-Sulfate in Epiphyses and Diaphyses of Suckling Rats

Dziewiatkowski

Ferrante

Bronner

et al. 1957

The Journal of Experimental Medicine

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The maximal accumulation of S 8~ in the epiphyseal cartilage of suckling rats occurs about 24 hours after the intraperitoneal injection of S~-sulfate (1), at which time the isotope is most highly concentrated in the epiphyseal cartilage plate (2). Thereafter, the concentration of S s6 in the epiphyses decreases rather slowly, the decrease being most noticeable in the epiphyseal cartilage plate and in regions of the epiphyses where ossification centers develop. In the diaphyses, on the other hand, there is a rapid rise and fall in the concentration of the isotope so that by the 4th hour only a faint autoradiographic reaction is elicited. Beginning at about the 24th hour the concentration of S 86 is seen to increase in the metaphyses. Because of the concurrent decrease of the concentration in the epiphyseal cartilage plate and increase in the metaphyses, it seemed likely that the S 8~ is transferred from the cartilage plates to the metaphyses, where it accumulates as inorganic sulfate or/and as some other compounds, of which the precursor is the chondroitin sulfate of the cartilage (2). Indeed, it has been reported that after the administration of S86-sulfate to dogs (3, 4), rabbits (5), and rats (6) the isotope in the bones can be separated into two forms, one of which probably is inorganic sulfate and the other a material akin to chondroitin sulfate.Experiments were undertaken to ascertain the nature, and the changes in concentration, of the S86-1abelled materials in epiphyses and diaphyses of suckling rats after administration of S3~-sulfate. The hope was that the findings would partially indicate the mechanisms involved in the development of the metaphyses. The results are presented here.EXPEI~ rM~.NTAL Experiment L--One hundred #c. of S 8~ as sodium sulfate I was injected intraperitoneally into each of twelve 7 day old rats, which were then returned to their mothers. Two animals

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“…D a t a for this clear association of the galactosamine with the carbohydrate were obtained by analyzing the hot formamide extracted carbohydrate and the mucopeptide residue for galactosamine by chromatographic column procedures described by Gardell (7). The results of analysis, given in Fig.…”

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confidence: 99%

Studies on the Chemical Structure of the Streptococcal Cell Wall

Krause

McCarty

1962

The Journal of Experimental Medicine

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In the preceding paper of this series, the trypsinized cell walls of Groups A and A-variant streptococci were shown to be composed of their respective group-specific polysaccharide antigen and a mucopeptide polymer (1). The data presented here indicate that Group C streptococcal cell walls contain the same two components. The mucopeptide is similar in composition to that of Group A streptococci with N-acetylglucosamine, N-acetylmuramic acid, and alanine, lysine, glutamic acid, and glycine as the principal constituents. The groupspecific polysaccharide also resembles that of Group A in that it is composed of rhamnose and hexosamine. However, in Group C polysaccharide the predominant hexosam~ne is N-acetylgalactosxmlne rather than N-acetylglucosamine. Previous studies have shown that the specificity of Group A carbohydrate is largely determined by terminal/3-N-acetylglucos~mln~de residues (2, 3) and in the present paper evidence is presented which indicates that N-acetylgalactosamine plays a comparable role in determining the serological specificity of group C carbohydrate. Materials and MetkodsThe following organisms were used for the preparation of cell walls and isolation of groupspecific carbohydrate: Group A, strain T12; Group A-variant, strain K43var.; Group C, strains H45A and 28RP95. A large quantity of strain H45A was kindly supplied by Lederle Laboratories. The methods for preparing cell walls and muralytic enzymes, and for extractions of carbohydrate both by enzymatic and chemical procedures have been previously described (1).Analytical Methods.--Analytical methods for rhamnose, glucosamine, muramic acid, and amino acids were the same as previously described (1).Because galactosamine as well as glucosamine is present in the cell wall of Group C streptococci, methods were required to estimate the relative amounts of these two sugars. For the preliminary work which required only an approximate determination of the relative amounts of glucosamine and galactosamine in a preparation the method of Tracey (4) was satisfactory. With this method a borate buffer is employed in the Elson-Morgan procedure which results

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Separation on Dowex 50 Ion-Exchange Resin of Glucosamine and Galactosamine and their Quantitative Determination.

Cited by 666 publications

References 0 publications

Distribution of sulphate and iduronic acid residues in heparin and heparan sulphate

Distribution of sulphate and iduronic acid residues in heparin and heparan sulphate

Turnover of S35-Sulfate in Epiphyses and Diaphyses of Suckling Rats

Studies on the Chemical Structure of the Streptococcal Cell Wall

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