Improved Method for Determination of Plasma Polysaccharides with Tryptophan.

Badin, J; Jackson, C. P.; Schubert, Maxwell

doi:10.3181/00379727-84-20620

Cited by 58 publications

(21 citation statements)

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“…of glucuronic acid developing color corresponding to 70 jug. mannose (25). Table II Figure 3 shows that the spectrum of the color produced by a mixture of glucuronic acid and lysozyme cannot be distinguished from that of the complex of lysozyme and the polysaccharide isolated from euglobulins.…”

Section: Methodsmentioning

confidence: 99%

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Plasma Polysaccharide Fraction Containing Uronic Acid, in Normal Subjects and in Patients With Rheumatoid Arthritis 1

Badin¹,

Schubert²,

Vouras³

1955

J. Clin. Invest.

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Section: Methodsmentioning

confidence: 99%

“…For determinations on polysaccharide-lysozyme complexes it was found important to have controls in the presence of a quantity of lysozyme similar to that in the precipitates. Figure 2 Total sugars determined with the modified tryptophane method (24,25) gave results with an error of about 10 per cent.…”

Section: Methodsmentioning

confidence: 99%

Plasma Polysaccharide Fraction Containing Uronic Acid, in Normal Subjects and in Patients With Rheumatoid Arthritis 1

Badin¹,

Schubert²,

Vouras³

1955

J. Clin. Invest.

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“…Alkali-soluble glucans were estimated from NaOH extraction supernatants by precipitating carbohydrates with 2 volumes of ethanol at Ϫ20ЊC. The total carbohydrate content of each fraction was measured as hexoses by the borosulfuric acid method (1). The data reported are the means of three experiments.…”

Section: Methodsmentioning

confidence: 99%

Increase in chitin as an essential response to defects in assembly of cell wall polymers in the ggp1delta mutant of Saccharomyces cerevisiae

et al. 1997

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The GGP1/GAS1 gene codes for a glycosylphosphatidylinositol-anchored plasma membrane glycoprotein of Saccharomyces cerevisiae. The ggp1⌬ mutant shows morphogenetic defects which suggest changes in the cell wall matrix. In this work, we have investigated cell wall glucan levels and the increase of chitin in ggp1⌬ mutant cells. In these cells, the level of alkali-insoluble 1,6-␤-D-glucan was found to be 50% of that of wild-type cells and was responsible for the observed decrease in the total alkali-insoluble glucan. Moreover, the ratio of alkali-soluble to alkali-insoluble glucan almost doubled, suggesting a change in glucan solubility. The increase of chitin in ggp1⌬ cells was found to be essential since the chs3⌬ ggp1⌬ mutations determined a severe reduction in the growth rate and in cell viability. Electron microscopy analysis showed the loss of the typical structure of yeast cell walls. Furthermore, in the chs3⌬ ggp1⌬ cells, the level of alkali-insoluble glucan was 57% of that of wild-type cells and the alkali-soluble/alkali-insoluble glucan ratio was doubled. We tested the effect of inhibition of chitin synthesis also by a different approach. The ggp1⌬ cells were treated with nikkomycin Z, a well-known inhibitor of chitin synthesis, and showed a hypersensitivity to this drug. In addition, studies of genetic interactions with genes related to the construction of the cell wall indicate a synthetic lethal effect of the ggp1⌬ kre6⌬ and the ggp1⌬ pkc1⌬ combined mutations. Our data point to an involvement of the GGP1 gene product in the cross-links between cell wall glucans (1,3-␤-D-glucans with 1,6-␤-D-glucans and with chitin). Chitin is essential to compensate for the defects due to the lack of Ggp1p. Moreover, the activities of Ggp1p and Chs3p are essential to the formation of the organized structure of the cell wall in vegetative cells.

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“…Analysis of the carbohydrate retained after dialysis gave the amount of alkali-insoluble (1->6)-fglucan. Carbohydrate was measured as hexose (17). (1--6)-3-Glucan was purified on a large scale for 'IC NMR analysis as described by Boone et al (28).…”

Section: Methodsmentioning

confidence: 99%

Yeast beta-glucan synthesis: KRE6 encodes a predicted type II membrane protein required for glucan synthesis in vivo and for glucan synthase activity in vitro.

Roemer

Bussey

1991

Proc. Natl. Acad. Sci. U.S.A.

134

138

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The KRE6 gene product is required for synthesis of the major 3glucans of the yeast cell wall, as mutations in this gene confer reduced levels of both the (1--6)-and (1-+3)-f3D-glucan polymers. Cloning ,-Glucan polysaccharides are widely found as cell surface polymers in bacteria, fungi, and plants. The biological role of these substances remains unclear, though they have been implicated in a range of processes, including providing osmotic and mechanical protection, selective permeation of macromolecules, cell extension, growth and division, and cell-cell interactions (1). In Saccharomyces cerevisiae, two classes of alkali-insoluble l3-glucans exist. (1-3)-3-Glucan, making up approximately 25% of the cell wall dry weight, has a degree of polymerization estimated as 1500 residues and is attached predominantly as 1-3 linkages and branched with occasional 1-*6 linkages (2).(1--6)-,-Glucan is a more complex, smaller, alkali-insoluble polymer with an average degree of polymerization of 140 residues and makes up approximately 7% of the cell wall dry weight (3). It is composed of a 1-*6-linked core, branched by occasional 1-+3 linkages, with 1-36 side chains providing many terminal glucopyranosyl residues (3). Despite the natural abundance of these polymers and the apparent simplicity of their structure as glucose homopolymers, little is known about the mechanism of 3-glucan biosynthesis. Recently the operon for cellulose synthesis in the bacterium Acetobacter xylinum has been analyzed, and four gene products, including two membrane proteins, have been shown to be required for the synthesis of this (1-*4)-f3-glucan (4, 5). In plants, the components of cellulose synthesis also appear to be membrane associated but have proved difficult to analyze biochemically. In synthesis of the plant (1--3)-f-glucan callose, a 57-kDa polypeptide has been implicated as a component of callose synthase by partial purification and photoaffinity labeling of this protein with an analog of the substrate UDP-glucose (6). In fungi (1-*3)-f-glucans are common cell wall components, and a considerable body of biochemical and physiological information exists on the (1-+3)-(3-glucan synthase (7). The synthase assayed in vitro is membrane associated, requires UDP-glucose as a substrate, and is stimulated by a soluble GTP-activated protein. Purification of these components has not been achieved. Unlike (1-*3)-1-glucan synthase, (1--6)-f-glucan synthase activity has not been detected in vitro. As with bacteria, genetics has been useful in identifying components in polysaccharide biosynthesis in fungi; examples include the N-linked glycosylation of proteins (8) and synthesis of chitin (9-11) and glycogen (12). However, the approach has been less informative for(1--3)-p3-glucan synthesis, where searches for conditional lethals with a lysis phenotype or for resistance to drugs thought to inhibit (1--3)-f-glucan synthesis have not identified synthase mutants. In yeast both chitin and glycogen synthesis use more than one synthase gene, and the res...

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Improved Method for Determination of Plasma Polysaccharides with Tryptophan.

Abstract: vacuum harvesting technic is described. The enumerated.

Cited by 58 publications

References 1 publication

Plasma Polysaccharide Fraction Containing Uronic Acid, in Normal Subjects and in Patients With Rheumatoid Arthritis 1

Plasma Polysaccharide Fraction Containing Uronic Acid, in Normal Subjects and in Patients With Rheumatoid Arthritis 1

Increase in chitin as an essential response to defects in assembly of cell wall polymers in the ggp1delta mutant of Saccharomyces cerevisiae

Yeast beta-glucan synthesis: KRE6 encodes a predicted type II membrane protein required for glucan synthesis in vivo and for glucan synthase activity in vitro.

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