Étude des structures fibrillaires de la sécrétion bronchique humaine

Havez, R; Roussel, P; Degand, Pierre; Biserte, G

doi:10.1016/0009-8981(67)90132-5

Cited by 46 publications

(7 citation statements)

References 21 publications

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“…3) and analysis of different sections of this peak indicates that the glycoprotein components show a continuous variation with respect to their sugar and sulphate content rather than a discrete population of different glycoproteins. This result differs from the observations of Havez and co-workers [42,44] that three separate groups of glycoproteins are present in sputum, namely neutral glycoproteins, sialoglycoproteins and sulphoglycoproteins. Whereas the present investigation does indicate that small amounts of neutral glycoprotein may be present it is difficult to eliminate the possibility that this did not arise as a result of the action of neuraminidase on a sialoglycoprotein since it is known that saliva, which is a contaminant of sputum, does contain an active neuraminidase [51].…”

Section: Discussioncontrasting

confidence: 99%

Isolation and Characterisation of Glycoproteins from Sputum

Roberts

1974

European Journal of Biochemistry

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Sputum has been separated into a sol and a gel phase by ultracentrifugation and the components of these phases examined. A glycoprotein of high molecular weight as well as serum albumin, immunoglobulin A, sll acid glycoprotein and lactoferrin were detected in the sol phase by gel filtration and immunodiffusion studies. A glycoprotein of similar chemical composition was isolated from the gel phase by solubilisation in 6 M urea/O. 1 M NaCl followed by fractional precipitation with ethanol.The carbohydrate content of this glycoprotein, termed the bronchial glycoprotein, varied from 58 (w/w) to 69% (w/w) with sputum from different patients and was composed of fucose, galactose, galactosamine, glucosamine and N-acetylneuraminic acid. Serine, threonine and proline constituted 44% (w/w) of the amino acid residues of the bronchial glycoprotein and an alkaline borohydride cleavage study indicated that 0-glycosidic linkages exist between N-acetylgalactosamine and the hydroxyamino acids in the peptide core. Fractionation of the bronchial glycoprotein on DEAESephadex A-25 yielded fractions enriched in sulphate and sialic acid but separate fuco, sialo and sulpho glycoproteins were not obtained. Similarly fractionation of the oligosaccharides released by the action of alkaline borohydride yielded a complex mixture of oligosaccharides but sulphated oligosaccharides free of sialic acid were not obtained. Separation of the oligosaccharides into two fractions of different sizes showed that sulphate content was greatest in the larger oligosaccharides whereas sialic acid content was highest in the smaller oligosaccharides. A methylation study of the bronchial glycoprotein, neuraminidase-digested bronchial glycoprotein and mild acid-hydrolysed bronchial glycoprotein revealed that the N-acetylneuraminic acid was linked to C-3 of penultimate galactose units. Fucose occurs in three different linkages, approximately 50 % is linked to C-2 of penultimate galactose units, approximately 25 % is linked to C-4 of glucosamine residues already substituted at

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Section: Discussioncontrasting

confidence: 99%

Isolation and Characterisation of Glycoproteins from Sputum

Roberts

1974

European Journal of Biochemistry

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“…Protease treatment of mucin. The naked peptide region of HTBM is more extensively degraded by pronase than by other proteases (8,22). Wells were coated with HTBM treated with pronase at 0.2 and 1.0 U of pronase (Calbiochem-Behring, San Diego, Calif.) per pLg of mucin in 2 mM calcium acetate buffer at 37°C for 24 h. HTBM in 2 mM calcium acetate buffer alone was used to coat control wells.…”

Section: Methodsmentioning

confidence: 99%

Tracheobronchial mucin receptor for Pseudomonas aeruginosa: predominance of amino sugars in binding sites

Vishwanath¹,

Ramphal²

1985

Infect Immun

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Pseudomonas aeruginosa, a common respiratory tract colonizer and pathogen, adheres to injured tracheal cells and to tracheobronchial mucin. These phenomena suggest that there are specific receptors for this organism in the respiratory tract. The receptor on injured tracheal cells contains n-acetylneuraminic acid as the principal sugar, but the structure of the receptor in mucin has not been described. Using a microtiter plate assay to study bacterial adherence to mucin, we have partially characterized the mucin receptor for P. aeruginosa. The receptor for both nonmucoid and mucoid strains is sensitive to periodate oxidation, suggesting that it is carbohydrate in nature, and the amino sugars n-acetylglucosamine and n-acetylneuraminic acid inhibited the adherence of both types of strains. Nonmucoid strains were more sensitive to inhibition by n-acetylneuraminic acid than to inhibition by n-acetylglucosamine, but the mucoid strains varied in their sensitivities to inhibition by each amino sugar. Preincubation of mucin with heat-inactivated influenza A virus (which binds to neuraminic acid) significantly reduced the adherence of P. aeruginosa. Treatment of mucin with Clostridium perfringens neuraminidase also reduced bacterial adherence significantly. Treatment of mucin with pronase did not affect adherence. Our results suggest that n-acetylglucosamine and n-acetylneuraminic acid are important constituents of the binding sites for P. aeruginosa on human tracheobronchial mucin.

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“…Gel viscosity was reduced with 0-075 M cysteine in 1/15 M phosphate buffer at pH 7-5, after which it was incubated at 370 C for two hours (Havez, Roussel, Degand, and Biserte, 1967). After dialysis at 40 C for five to seven days, the specimen was centrifuged at 15,000 rev/min for 30 minutes, and the supernatant was used for further testing.…”

Section: Methodsmentioning

confidence: 99%

Effect of disodium cromoglycate (Intal) on sputum protein composition

Heilpern¹,

Rebuck²

1972

Thorax

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The protein content in the sputum of nine patients with asthma was compared with that in 11 non-asthmatic patients by a method of gel electrophoresis. It was found that the albumin content was significantly higher in the sputum of the asthmatics than of the non-asthmatics (P<001). A further 11 asthmatic patients who were taking disodium cromoglycate (Intal) as part of their treatment had sputum albumin levels indistinguishable from the non-asthmatics. When patients with asthma were then studied serially, before and after disodium cromoglycate therapy, the albumin content returned to non-asthmatic levels within two days of starting treatment. It is suggested that disodium cromoglycate has a previously unrecognized action in altering sputum proteins.Methods have now been developed for investigating the biochemical characteristics of bronchial mucus (Ryley and Brogan, 1968) as well as sputum viscosity (Cobbin, Elliott, and Rebuck, 1971) in patients with asthma and chronic lung disease.Sputum production in asthma is markedly decreased in patients treated with disodium cromoglycate (Intal). This drug prevents asthmatic attacks attributable to precipitins (Pepys, Hargreave, Chan, and McCarthy, 1968) as well as having a steroid-sparing effect (Read and Rebuck, 1969).Although various proteins in the sputum of patients with asthma have been studied, and compared with those of normal subjects (Newcomb and DeVald, 1969;Dennis, Hornbrook, and Ishizaka, 1964) GROUP D These five patients, all female, had asthma, and were tested both before and after starting disodium cromoglycate therapy, so that they were able to act as their own controls. Their ages ranged from 17 to 46 (mean 34) years.In no case was bloodstained sputum studied, and patients whose sputum showed bacteriological evidence of pathogenic infection were eliminated from the trial.Sputum samples consisted of the total sputum produced by patients during the first four hours after waking. The samples were expectorated directly into jars containing 10 ml tris-glycine buffer at pH 8-3.The gel fraction was separated by straining the sputum through gauze for 30 minutes. Gel was then scraped from the gauze and its volume was measured.Gel viscosity was reduced with 0-075 M cysteine in 1/15 M phosphate buffer at pH 7-5, after which it was incubated at 370 C for two hours (Havez, Roussel, Degand, and Biserte, 1967). After dialysis at 40 C for five to seven days, the specimen was centrifuged at 15,000 rev/min for 30 minutes, and the supernatant was used for further testing. 726 on 10 May 2018 by guest. Protected by copyright.

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Étude des structures fibrillaires de la sécrétion bronchique humaine

Cited by 46 publications

References 21 publications

Isolation and Characterisation of Glycoproteins from Sputum

Isolation and Characterisation of Glycoproteins from Sputum

Tracheobronchial mucin receptor for Pseudomonas aeruginosa: predominance of amino sugars in binding sites

Effect of disodium cromoglycate (Intal) on sputum protein composition

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