Qualitative and quantitative analysis of reducing carbohydrates by radiochromatography on ion-exchange papers

Conrad, H. Edward; Varboncouer, S.R. Evelyn; James, Matthew E.

doi:10.1016/0003-2697(73)90505-8

Cited by 55 publications

(15 citation statements)

References 11 publications

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“…The purified disaccharide was hydrolyzed to its monosaccharide units by incubation in 70% formic acid overnight at 100°C. To analyze the constituent monosaccharides, the hydrolyzed sample was reduced with sodium [ 3 H]borohydride (6) to label all reducing sugars. The identity of the reduced sugars was determined by descending paper chromatography with Whatman no.…”

Section: Chemicalsmentioning

confidence: 99%

2-Sulfotrehalose, a novel osmolyte in haloalkaliphilic archaea

et al. 1997

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A novel 131 ␣-linked glucose disaccharide with sulfate at C-2 of one of the glucose moieties, 1-(2-O-sulfo-␣-D-glucopyranosyl)-␣-D-glycopyranose, was found to be the major organic solute accumulated by a Natronococcus sp. and several Natronobacterium species. The concentration of this novel disaccharide, termed sulfotrehalose, increased with increasing concentrations of external NaCl, behavior consistent with its identity as an osmolyte. A variety of noncharged disaccharides (trehalose, sucrose, cellobiose, and maltose) were added to the growth medium to see if they could suppress synthesis and accumulation of sulfotrehalose. Sucrose was the most effective in suppressing biosynthesis and accumulation of sulfotrehalose, with levels as low as 0.1 mM being able to significantly replace the novel charged osmolyte. Other common osmolytes (glycine betaine, glutamate, and proline) were not accumulated or used for osmotic balance in place of the sulfotrehalose by the halophilic archaeons.

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Section: Chemicalsmentioning

confidence: 99%

2-Sulfotrehalose, a novel osmolyte in haloalkaliphilic archaea

et al. 1997

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“…Part of the initial evidence implicating 6-phos- 7 8 THE JOURNAL OF CELL BIOLOGY " VOLUME 84,1980 phomannose recognition in the uptake of acid hydrolases by fibroblasts was the competitive inhibition of uptake by phosphomannans containing 6-phosphomannose (13) . We subsequently reported on the correlation of structural features of phosphomannans and phosphomannan hydrolysis fragments with their ability to inhibit enzyme pinocytosis (15) .…”

Section: Abstract Receptorlysosomal Enzymes Pinocytosis -Multivalencmentioning

confidence: 99%

Fibroblast receptor for lysosomal enzymes mediates pinocytosis of multivalent phosphomannan fragment

Hd¹,

Natowicz²,

Sly³

et al. 1980

The Journal of Cell Biology

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Mild acid hydrolysis of phosphomannan secreted by the yeast hansenula holstii (NRRL Y- 2448) produces two phosphomannyl fragments which differ strikingly in their potency as inhibitors of pinocytosis of human β-glucuronidase by human fibroblasts. The larger molecular weight polyphosphomonoester fragment is 100,000-fold more potent an inhibitor of enzyme uptake than the smaller penta-mannosyl-monophosphate fragment. Binding to attached fibroblasts at 3 degrees C was much greater with the polyphosphomonoester fragment than with the pentamannosyl-monophosphate. The larger molecular weight fragment was also subject to adsorptive pinocytosis and was taken up by fibroblasts at a rate 30- fold greater than the rate of uptake of pentamannosyl-monophosphate. Evidence that the polyphosphomonoester fragment is taken up by the phosphomannosyl-recognition system that mediates uptake of lysosomal enzymes includes: (a) its pinocytosis is inhibited by the same compounds that competitively inhibit enzyme pinocytosis (mannose-6-phosphate and phosphomannan from saccharomyces cerevisiae mutant mnn-1); (b) alkaline phosphatase treatment greatly reduces its susceptibility to pinocytosis; (c) its pinocytosis is competitively inhibited by high-uptake human β-glucuronidase; and (d) this inhibition by high-uptake enzyme is dramatically reduced by prior treatment of the enzyme with alkaline phosphatase or endoglycosidase-H. Endoglycosidase-H treatment human β-glucuronidase dramatically reduced its susceptibility to pinocytosis by fibroblasts. The phosphomannosyl components of high- uptake enzyme released by endoglycosidase-H treatment were much less effective inhibitors of polyphosphomonoester pinocytosis than when present on the phosphomannyl-enzyme. These results suggest that high-uptake acid hydrolases may be polyvalent ligands analogous to the polyphosphomonoester mannan fragment whose pinocytosis depends on interaction of more than one phospho-mannosyl recognition marker with pinocytosis receptors on fibroblasts.

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“…In fact, it is desirable to dialyze the glycosaminoglycan solution to remove all salts before beginning the cleavage step. Another reason for the pre-dialysis is that the NaB[ 3 H] 4 reagent is catalytically destroyed by oxyanions, such as PO 4 3− (Conrad et al, 1973).…”

Section: Critical Parametersmentioning

confidence: 99%

Nitrous Acid Degradation of Glycosaminoglycans

Conrad

1995

CP Molecular Biology

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Glycosaminoglycans (GAGs) are made up of disaccharide units that are distinguished from each other by the monosaccharide units of which they are composed and by the degree and position of sulfation. These disaccharide units represent the monomeric units of the GAG; thus, measurement of the disaccharide composition of a GAG represents the first step in the characterization of the polymer. In this unit, cleavage of the glycosidic bonds of the N-sulfated GlcN residues in heparin and heparan sulfate is described, in addition to cleavage of the bonds between the N-acetylated amino sugar residues in heparin, heparan sulfate, chondroitin sulfate, dermatan sulfate, and keratan sulfate, and hyaluronic acid. Using these procedures involving, all GAGs can be converted completely to their constituent disaccharides and reduced with NaB[3H]4) to yield labeled disaccharides that can be assayed qualitatively or quantitatively. The procedure may also be used to analyze metabolically labeled GAGs (with or without the use of NaB[3H]4).

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Qualitative and quantitative analysis of reducing carbohydrates by radiochromatography on ion-exchange papers

Cited by 55 publications

References 11 publications

2-Sulfotrehalose, a novel osmolyte in haloalkaliphilic archaea

2-Sulfotrehalose, a novel osmolyte in haloalkaliphilic archaea

Fibroblast receptor for lysosomal enzymes mediates pinocytosis of multivalent phosphomannan fragment

Nitrous Acid Degradation of Glycosaminoglycans

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