Purification et études des mucines de kystes bronchogéniques

Degand, Pierre; Roussel, Philippe; Lamblin, Geneviève; Havez, R

doi:10.1016/0304-4165(73)90312-7

Cited by 31 publications

(10 citation statements)

References 14 publications

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“…Prior to staining with Alcian blue, gels were fixed as described by Kunicki et al [13]. Agarose gel electrophoresis was carried out on 0.9% agarose slabs (7 x 2.5 cm), in 0.1 M barbital buffer, pH 8.2 [14], followed by staining with amidoschwarz, toluidine blue and the periodic acid/Schiff reaction. Crossed immunoelectrophoresis was performed as previously described [S].…”

Section: Electrophoresismentioning

confidence: 99%

The major human urinary trypsin inhibitor is a proteoglycan

et al. 1986

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The major urinary trypsin inhibitor (MI 44000), isolated from human urine, contains 35% carbohydrate. In addition to N-acetylglucosamine and neutral sugars (primarily mannose and galactose), the carbohydrate moiety contains hexuronic acid and N-acetylgalactosamine and corresponds to a glycosaminoglycan. This carbohydrate chain is an integral component of the inhibitor: it does not dissociate from the inhibitor when using dissociative conditions such as sodium dodecyl sulfate, guanidinium chloride, or by increasing ionic strength or mixing with cetylpyridinium chloride.This glycosaminoglycan chain is sensitive to chondroitinase ABC or testicular hyaluronidase digestion and corresponds to slightly sulfated chondroitin 4-sulfate or 6-sulfate. After treatment by these enzymes, the urinary inhibitor has a lower molecular mass (MI 26000) but still inhibits trypsin.Several acid-resistant trypsin inhibitors with molecular masses, estimated by sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE), ranging from 40 kDa -45 kDa, have been isolated from human urine [l -41. All these inhibitors show an immunological cross-reactivity with intera-trypsin inhibitor.Hochstrasser et al.[5] first described a physiological urinary inhibitor having an apparent molecular mass of 34 kDa as calculated from its specific activity. This inhibitor (HI-30) has an apparent molecular mass of 30 kDa, as estimated by gel chromatography [6], but 40 kDa in SDS-PAGE [4].We previously described the isolation and characterization of a human urinary trypsin inhibitor (UTI) with the same aminoterminal sequence as 81. This sequence was recently found to be identical with that of inter-a-trypsin inhibitor [9, 101; those data tend to confirm the hypothesis that the different urinary trypsin inhibitors isolated so far are derived from inter-a-trypsin inhibitor.The proportion and nature of the carbohydrate reported for those inhibitors is variable, ranging from 10% to 50% [l,3, 71. According to Hochstrasser et al. [ll] the carbohydrate content of HI-30 accounts for 50% of the total weight and includes two carbohydrate chains: one is attached N-glycosidically via an asparaginyl residue, the other is 0-glycosidically linked to the polypeptide via a seryl residue in position 10. This last chain is made up of at least 48 Nacetylgalactosamine, 2 glucose, 18 galactose and 2 N-acetylneuraminic acid residues [6]. Such a composition is quite unusual for a glycoprotein.In this report we describe a slightly modified procedure for isolation of urinary trypsin inhibitor and we present evidence that it consists a proteoglycan. MATERIALS AND METHODSSephacryl S-200, DEAE-Sephacel and ConA-Sepharose were purchased from Pharmacia, DEAE-Trisacryl was from IBF. Acrylamide and N-N'-methylenebisacrylamide were from BDH, agarose (Indubiose A 37) from IBF. Bovine pancreatic trypsin (treated with Tos-Phe-CH2C1) and L-benzoylarginine p-nitroanilide were purchased from Merck, chondroitinase ABC (from Proteus vulgaris) and bovine testicular hyaluronidase were from ...

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Section: Electrophoresismentioning

confidence: 99%

The major human urinary trypsin inhibitor is a proteoglycan

et al. 1986

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“…Baker et al (1973) have demonstrated an increase in glycosyltransferases from microsomal fractions prepared from respiratory tract tissue showing goblet cell hyperplasia and gland hyperplasia, from dogs exposed to sulphur dioxide. Degand et al (1973) have suggested that an increase in galactosamine leads to an increase in the length of the glycan chains and thus to a possible increase in the number of sites available for sulphation of the glycoprotein. It may be that the increase in sulphomucin seen in the cells of the hypertrophied pig bronchial gland is secondary to synthesis of galactosamine.…”

Section: Jones Baskerv~lle and Reidmentioning

confidence: 99%

Histochemical identification of glycoproteins in pig bronchial epithelium: (a) normal and (b) hypertrophied from enzootic pneumonia

Jones

Baskerville

Reid

1975

The Journal of Pathology

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The glycoproteins in the normal pig bronchial gland are identified by the combined Alcian Blue (AB)-periodic acid Schiff (PAS) technique, with the use of sialidase digestion and AB staining either at pH 2-6 or at pH 1-0. In enzootic pneumonia (produced experimentally by infection with Mycoplasma hyorhinis) the bronchial gland hypertrophies, mucous and serous cells both increase, in number and size; hence the total glycoprotein content of the gland increases. The distribution of glycoproteins in the hypertrophied gland differs from that in the normal. Quantitative analysis of the mucous cells shows that in the hypertrophied gland the acid glycoprotein is increased relative to the neutral. There is also a relative change in the amounts of sialidase-sensitive sialomucin and sulphomucin; both are significantly increased at the expense of the sialidase-resistant sialomucin. Qualitative analysis of the serous cells shows that in the normal gland most of the glycoprotein is neutral and that the small amount of acid glycoprotein is sialidase-resistant sialomucin. In the hypertrophied gland there is relatively more acid glycoprotein which is either sialidase-resistant sialomucin or sulphomucin; in addition, in pigs with enzootic pneumonia there is an increase in the height of the bronchial epithelium and a depletion in both goblet cell number and glycoprotein content, which latter has more neutral glycoprotein and less acid glycoprotein.

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“…Polyclonal antiserum raised against tracheobronchial apomucins Tracheobronchial mucins from human bronchial sputum were prepared and purified as in [10,18]. An antigenic mixture made of a pool of deglycosylated peptides allowed the preparation of an anti-airway apomucin antiserum [10,19].…”

Section: Methodsmentioning

confidence: 99%

Characterization of the human mucin gene MUC5AC: a consensus cysteine-rich domain for 11p15 mucin genes?

Duperat

Audie

Debailleul

et al. 1995

Biochemical Journal

193

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To date five human mucin cDNAs (MUC2, 5A, 5B, 5C and 6) mapped to 11p15.3-15.5, so it appears that this chromosome region might contain several distinct gene loci for mucins. Three of these cDNAs, MUC5A, B and C, were cloned in our laboratory and previously published. A common number, 5, was recommended by the Human Gene Mapping Nomenclature Committee to designate them because of their common provenance from human tracheobronchial mucosa. In order to define whether they are products of the same gene locus or distinct loci, we describe in this paper physical mapping of these cDNAs using the strategy of analysis of CpG islands by pulse-field gel electrophoresis. The data suggest that MUC5A and MUC5C are part of the same gene (called MUC5AC) which is distinct from MUC5B. In the second part of this work, complete sequences of the inserts corresponding to previously described (JER47, JER58) and novel (JER62, JUL32, MAR2, MAR10 and MAR11) cDNAs of the so-called MUC5AC gene are presented and analysed. The data show that in this mucin gene, the tandem repeat domain is interrupted several times with a subdomain encoding a 130 amino acid cysteine-rich peptide in which the TR3A and TR3B peptides previously isolated by Rose et al. [Rose, Kaufman and Martin (1989) J. Biol. Chem., 264, 8193-8199] from airway mucins are found. A consensus peptide sequence for these subdomains involving invariant positions of most of the cysteines is proposed. The consensus nucleotide sequence of this subdomain is also found in the MUC2 gene and in the MUC5B gene, two other mucin genes mapped to 11p15. The functional significance for secreted mucins of these cysteine-rich subdomains and the modular organization of mucin peptides are discussed.

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Purification et études des mucines de kystes bronchogéniques

Cited by 31 publications

References 14 publications

The major human urinary trypsin inhibitor is a proteoglycan

The major human urinary trypsin inhibitor is a proteoglycan

Histochemical identification of glycoproteins in pig bronchial epithelium: (a) normal and (b) hypertrophied from enzootic pneumonia

Characterization of the human mucin gene MUC5AC: a consensus cysteine-rich domain for 11p15 mucin genes?

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