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Query: UMLS:C0272170 (SDS)
 50,377 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Ocular mucin, the major product of conjunctival goblet cells, constitutes the innermost layer of preocular tear film. Ocular mucin is known for its limited amount and inaccessibility. Using impression cytology, mucus strands collected from the inferior fornix of either rabbit or human eyes were found to contain inflammatory cellular debris. In order to circumvent these difficulties and to isolate native mucin molecule(s), we bathed rabbit eyes in fluid containing isotonic PBS and 5.5 X 10(-4) M acetylcholine for 4 or 12 hr. Bathing fluids containing rabbit ocular mucin (ROM), 1 ml per eye, were pooled and combined with 1M guanidine HCl and protease inhibitors containing EDTA, PMSF, and sodium azide to avoid any possible enzymatic degradation, and then separated under the same conditions by Sepharose CL-4B. In parallel, commercial porcine stomach mucin (PSM) was purified and used to compare with ROM. We also developed nitrocellulose-based dot semi-quantitative assays for nucleic acid, protein, and glycoprotein. PAS-positive fractions monitored by such a dot assay were collected at CL-4B void volume and then separated from nucleic acid contaminants by CsCl-gradient ultracentrifugation. A protein fraction, 65K, poorly-glycosylated, with high contents of Asx, Glx, and Gly was found strongly associated with both ROM and PSM, and was only separable by ultracentrifugation in 4M guanidine HCl and CsCl. Purification of the ROM was verified by SDS-polyacrylamide gel electrophoresis, amino acid analysis, and carbohydrate analysis. These results will allow future exploration of the molecular mechanism by which tear film is achieved.
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PMID:Purification and characterization of rabbit ocular mucin. 362 33

Using gastric mucous cells which are involved exclusively in the synthesis of secretory O-glycosidic glycoprotein (mucin), the relationship between protein core synthesis and its acylation with fatty acids was investigated. Labeling of the cells with [3H]palmitic acid and [35S]methionine followed by isolation of peptidyl-tRNA and release of nascent peptides, indicated that these peptides contain covalently bound fatty acids. The high performance thin layer chromatography, SDS-gel electrophoresis, and radioactivity scanning revealed that the preparation contained three fractions labeled with palmitate (Mr 15,000-3,600) and two (Mr 1,500 and less) without this label. Based on these data and the nascent peptides amino acid analysis, we conclude that the protein core of the O-glycosidic glycoprotein is acylated with fatty acids during translation, when the peptide chain is longer than 21 amino acid residues.
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PMID:Cotranslational attachment of fatty acids to nascent peptides in gastric mucus glycoprotein. 364 23

Individual mucus samples were collected from normal individuals and from patients with ocular cicatricial pemphigoid (CP), Stevens-Johnson syndrome (SJS), and various types of conjunctival inflammation (rosacea, meibomianitis, atopy, keratoconjunctivitis sicca, etc.). The mucus samples were dissolved in sample buffer containing 8M urea, 2% SDS and 5% 2-mercaptoethanol and were electrophoresed on gradient 2-16% polyacrylamide gels. Four glycoproteins with molecular weights greater than 200,000 daltons were consistently observed in both individuals with normal conjunctiva and patients with CP, SJS, and other diseases exhibiting conjunctival inflammation. The amounts of each glycoprotein appeared to vary from one individual to another; however, the presence or absence of specific glycoproteins could not be correlated with the different ocular diseases. The techniques described for mucus analysis offer advantages over previously published techniques since improved resolution of the mucous glycoproteins can be achieved by electrophoresis on 2-16% gradient gels, and individual samples can be analyzed. Our results suggest that substantial amounts of ocular mucous glycoprotein are present in the eyes of patients with CP and SJS, diseases which have been previously described as mucin-deficient dry eye syndromes.
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PMID:SDS-gradient polyacrylamide gel electrophoresis of individual ocular mucus samples from patients with normal and diseased conjunctiva. 378 Feb 81

A lectin purified from the Tora-bean (Phaseolus vulgaris) by affinity chromatography with Con-A Sepharose was shown to be a glycoprotein containing 7.8% neutral sugars (D-mannose, N-acetyl-D-glucosamine, L-fucose, and D-xylose, in a molar ratio of 9.6 : 2.0 : 0.6 : 0.7). Its molecular weight was 130,000, as estimated by exclusion gel chromatography, and SDS gel electrophoresis showed that it consists of four subunits of molecular weight 32,000. The lectin reacts with various glycoproteins, i.e., blood group substances, human parotid salivary glycoprotein, fetuin, and bovine submaxillary mucin. Divalent cations, such as Ca2+, Mn2+, and Mg2+, appear to stimulate its reactivity. Inhibition tests using the glycopeptide fragment from fetuin and some oligosaccharides, as well as the binding test with 14C-N-acetyl-lactosamine suggest that the sequence of D-galactose, N-acetyl-D-glucosamine, and D-mannose residues in the carbohydrate chain of fetuin is essential for binding.
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PMID:Purification and characterization of Tora-bean (Phaseolus vulgaris) lectin. 615 49

The protein in palatine saliva was investigated and compared with that in parotid saliva by means of SDS electrophoretic method. It was found that palatine saliva contains a large amount of high molecular weight protein (higher than 300,000) which is identified as mucin-rich glycoprotein and sialoglycoprotein, while no such protein is present in parotid saliva. The protein composition of palatine saliva has qualitatively smaller individual difference when compared to that of parotid saliva. We suggest that the high molecular weight protein which is specific to palatine saliva probably contributes to the rentention of complete maxillary dentures.
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PMID:Electrophoretic analysis of the protein in palatine saliva. 615 20

We report, for the first time, immune responses within two lines of inbred rats to a purified Lewis rat glycoprotein antigen which is organ-specific for intestine. The antigen was prepared by solubilization of gut epithelial cell-associated macromolecules, fractionation in ethanol, and molecular sieve chromatography over Sepharose 2B. Homogeneity of the end product (RGCG-PK1) was supported by results of both double diffusion in agar and SDS polyacrylamide gel electrophoresis. Amino acid analysis and specific sugar determination proved that RGCG-PK1 was not a classical mucin because of its comparatively high tyrosine and low galactosamine + glucosamine content, and the absence of glycosidic linkages to serine and threonine. Organ-specificity was shown by the ability of RGCG (but not liver homogenate) to inhibit precipitation and haemagglutination by heterologous specific sera. Organ-specificity was confirmed by the demonstration of RGCG-PK1-specific immunofluorescence staining of rat small and large intestine, but not esophagus, stomach or liver. RGCG-PK1 determinants within rat and human small bowel were found to be confined to goblet cells and intestinal glycocalyx. Anti-RCGC-PK1 serum showed no reactivity with highly purified xenogeneic mucins nor with syngeneic small bowel mucin. Specific antibody (as well as antibody-dependent cellular cytotoxicity) to RGCG was elicited and detected for up to 10 days in two lines of inbred rats, including the one (Lewis) from which the antigen was isolated. The duration and peak of the humoral immune response were abbreviated compared with that of a xenogeneic control glycoprotein studied in parallel, probably due to immunoregulatory mechanisms operative for self antigens.
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PMID:Goblet cell glycoprotein: an organ-specific antigen for gut. Isolation, tissue localization and immune response. 617 74

Previous studies have shown that human small-intestinal mucin consists of high-Mr glycoproteins and a smaller S-S-bonded protein of 118 kDa. The major antigenic determinants of the mucin were associated with the large glycoproteins, but depended for stability on intact disulphide bonds, and were destroyed by digestion with Pronase. In the present study we isolated and analysed the component parts of mucin from patients with cystic fibrosis with special attention being paid to the peptide constituents. After reduction with 0.2 M-beta-mercaptoethanol [5 min, 100 degrees C in 1% SDS (sodium dodecyl sulphate)], the large glycoproteins and smaller peptide with an apparent molecular size of 118 kDa were separated by equilibrium density-gradient centrifugation in CsCl, Sepharose 4B chromatography or preparative SDS/polyacrylamide-gel electrophoresis. The large glycoproteins contained about 70% of the protein of the native mucin. Digestion with Pronase resulted in a further loss of 'naked' protein (10% of the native mucin protein) from the C-terminal end of the glycoprotein peptide core, and left behind highly glycosylated proteins comprised mainly (70 mol%) of threonine, serine and proline. The 118 kDa component, which contained about 30% of the native mucin protein, consisted mainly of aspartic acid, serine, glutamic acid and glycine (40 mol%), plus threonine, proline, alanine, valine and leucine (35 mol%). Together with the 'naked' protein segment, the 118 kDa component contained most of the cysteine residues of the native mucin. Surprisingly, the peptide also contained carbohydrate (less than or equal to 5% of the native mucin carbohydrate but 50% by weight of the 118 kDa component), which included 9 mol% mannose, suggesting the presence of N-linked oligosaccharides. The peptide exhibited strong non-covalent interactions with the high-Mr glycoproteins and a tendency to self-aggregate in the absence of dissociating agents. Our findings therefore suggest that native mucin consists of large glycoproteins capable of forming disulphide bridges from their C-terminal 'naked' (antigenic) regions to a smaller glycopeptide having an Mr of 118 000.
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PMID:Biochemical characterization of the component parts of intestinal mucin from patients with cystic fibrosis. 651 57

Immunodiffusion, immunoelectrophoresis, SDS-polyacrylamide gel electrophoresis, polyacrylamide gel electrophoresis, ultracentrifugation analysis, and gel permeation chromatography in chaotropic and detergent mediums showed that the human seminal plasma obtained and stored in the usual way is mainly composed by heterodispersed glycoproteins of low molecular weight. The glycoprotein components of the human seminal plasma which do not interact specifically with concanavalin A (con A) were separated by column chromatography on Con A-Sepharose according to nonspecific ion exchange and hydrophobic interactions with the protein. The carbohydrate composition of the glycoproteins and the existence of the N-acetylgalactosaminylserine (or threonine) linkage, as well as their aggregation properties show that they are mucin-type glycoproteins. They resulted "immunologically pure" although the carbohydrate chains show the structural heterodispersion usually found in mucins. The glycoproteins have a common structural pattern for the terminal nonreducing, determinant-bearing sequences of their oligosaccharide chains which, together with the low molecular weights and the known fact that mucus secretions of normal cells contain only one or two types of glycoproteins, suggest that they are fragments produced by endogenous proteolytic degradation of a native mucin.
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PMID:The antigenic mucin-like glycoproteins of human seminal plasma. 665 Jul 11

We report here a novel genetically determined polymorphism of a human urinary mucin which is demonstrable by the separation technique of SDS polyacrylamide gel electrophoresis, followed by detection with radio-iodinated lectins. The mucins are demonstrable using various lectins but the polymorphism is most easily recognized using peanut agglutinin and we therefore propose to designate this new genetic locus PUM (peanut-reactive urinary mucin). Four common alleles have been identified and an autosomal codominant mode of inheritance has been found in the families studied so far.
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PMID:A genetic polymorphism of a human urinary mucin. 665 Dec 17

Milk fat globule membrane (MFGM) enclosing fat droplets in human milk was found to contain a high molecular weight glycoprotein which did not migrate in 10% acrylamide gel on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This glycoprotein (termed PAS-0) was isolated by Sepharose CL-4B chromatography. Isolated PAS-0 gave one band on SDS-PAGE using 5% acrylamide gel (acrylamide : bisacrylamide = 4 : 1, w/w) and gave one peak on analytical ultracentrifugation, indicating its homogeneity. PAS-0 was rich in serine, threonine, proline, glycine, and alanine. In contrast, contents of sulfur-containing amino acids were very low. Fucose, galactose, N-acetylglucosamine, N-acetylgalactosamine, and sialic acid were detected as constituent sugars of PAS-0 and the total carbohydrate content was about 50%. Alkali-borohydride treatment suggested that the carbohydrate moiety was linked to the polypeptide core with O-glycosidic bond(s). There results suggested that PAS-0 was a mucin-like glycoprotein. PAS-0 was shown to be resistant to pepsin, trypsin and chymotrypsin digestion, but susceptible to Pronase and Subtilisin BPN'. Extraction of intact milk fat globules (cream) with MgCl2 and guanidine hydrochloride solutions suggested that PAS-0 was an intrinsic component of MFGM. Digestion of cream with Subtilisin BPN' demonstrated that PAS-0 was located on the external surface of fat globules and was accessible to molecules outside the globules. By agglutination-inhibition tests using eight lectins, PAS-0 was suggested to act as surface receptors for Ricinus communis agglutinin, wheat germ agglutinin and peanut agglutinin.
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PMID:Isolation and characterization of mucin-like glycoprotein in human milk fat globule membrane. 706 73


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