Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.21.4 (trypsin)
 42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Heparitinase [EC 4.2.2.8, heparitin sulfate lyase] was prepared from an extract of cultured cells of Flavobacterium heparinum. Purification of the enzyme was achieved by repeating the hydroxyapatite column chromatography. The enzyme was used to degrade heparan sulfate occurring on the surfaces of ascites hepatoma cells, AH 66. From the supernatant of the enzyme-treated cells, breakdown products from heparan sulfate could be detected by paper chromatography. The heparitinase was found to be more effective than trypsin in removing heparan sulfate from the cells. Furthermore, on analyzing glycosaminoglycans and glycopeptides from the enzyme-treated cells and control cells, it was concluded that heparan sulfate was exclusively present on the cell surface and accessible to the heparitinase whereas other cell surface complex carbohydrates remained intact.
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PMID:Degradative removal of heparan sulfate from the surface of an ascites hepatoma, AH 66, by heparitinase. 645 48

Cultured bovine corneal endothelial cells express 5-8 ng basic fibroblast growth factor (bFGF)/mg cell protein and distribute it between the intracellular and pericellular compartment. Confluent cultures retain approximately 80% of the total bFGF intracellularly, whereas 20% is present in the pericellular (trypsin-releasable) compartment. No bFGF can be detected in the culture medium. The presence of 1-2 ng/ml medium of endogenous or exogenous (human recombinant) bFGF is sufficient to support cell growth. Simultaneously, cells incorporate [35S]sulfate and [3H]glucosamine into the sulfated proteoglycans associated with the cell layer at a rate that is three times higher than in the absence of bFGF. The enhanced proteoglycan synthesis is accompanied by a shift in proteoglycan distribution. In control cells, cell-associated heparan sulfate accounts for about 30% of the total glycosaminoglycans, whereas under the influence of bFGF the amount of heparan sulfate increases to approximately 60%. At the same time, the molecular structure of the heparan sulfate molecule undergoes bFGF-specific changes as indicated by the [35S]oligosaccharide pattern generated by heparitinase I degradation. The proportion of [35S]oligosaccharides with greater than six monosaccharides decreases on account of disaccharides and tetrasaccharides under the influence of bFGF. Pretreatment of bFGF with neutralizing antibodies against bFGF abolishes its biological activity. The results suggest a bFGF-dependent change in the rate of synthesis and structural features of the membrane-associated heparan sulfate in corneal endothelial cells. The modification of the heparan sulfate structure could influence its bFGF-binding and antiproliferative activity.
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PMID:Basic fibroblast growth factor controls the expression and molecular structure of heparan sulfate in corneal endothelial cells. 853 92

When cells dissociated from the neonatal rat brains are plated on a poly-lysine-coated surface in a serum-free medium, they display a strange morphology: a dark and extended cell body. Preincubation of the surface with fetal bovine serum was found to inhibit the appearance of this strange contraction of the basal cell sheets in a dose-dependent manner. This finding indicated the presence of a factor(s) in the serum, which might be an appropriate substratum for prolonged survival of brain neurons. In the current study, this factor was highly purified through DEAE ion-exchange chromatography followed by gel filtration. The factor was eluted from a Superose column at fractions corresponding to a molecular weight greater than 1000 kDa. By SDS-PAGE analysis, these fractions were found to contain a major band (>/=1000 kDa) positive for alcian blue and few minor bands faintly stainable with Coomassie blue. The activity of the purified sample, inducing the morphological change in cells, was diminished by incubation with chondroitinase ABC. Neither heparitinase II, hyaluronidase, nor trypsin modified the activity. An authentic chondroitin sulfate (type B) mimicked the serum action on the morphology of brain cells in early stages of culture. Taking these findings together, it is suggested that the factor in serum beneficial for the attachment of brain cells is composed of a chondroitin sulfate with a Mr greater than 1000 kDa. Cortical cells dissociated from the neonatal rat brain attached well to the purified factor-coated surface and displayed a healthy morphology: an optically-reflective cell body with thick neurites for at least 3 days in the absence of serum.
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PMID:A culture substratum appropriate for brain cells is a chondroitin sulfate glycosaminoglycan in serum. 947 1

Tight junctions seal polarised surface epithelial respiratory cells so as to prevent the passage of bacteria and toxins through the epithelial sheet. Disruption of tight junctions, which may occur during injury and repair processes of airway epithelium, favours potential bacterial interaction with receptors from cell basolateral membranes. Earlier studies reported that non-polarised and untight epithelial respiratory cells are highly susceptible to Pseudomonas aeruginosa adherence and internalisation. As heparan sulphate proteoglycans (HSP) from cell basolateral membranes in epithelial cells without tight junctions may become accessible to bacterial ligands, the present study investigated their role as potential receptors for non-piliate P. aeruginosa ligands. Treatment of cells with heparitinase I and II significantly reduced (51.2% and 51.7%, respectively) P. aeruginosa adherence to epithelial respiratory cells without tight junctions. The internalisation of bacteria was not affected by treatment with heparitinases. Treatment of the bacteria with heparin and heparan sulphate also significantly reduced their adherence to respiratory cells (34.3% and 43.7%, respectively). Treatment of cells with other enzymes (trypsin, lipase and chondroitinase ABC) or treatment of bacteria with chondroitin-4-sulphate did not modify the adherence to respiratory cells significantly. Both affinity chromatography and Western blotting assays showed the interaction of different P. aeruginosa outer-membrane proteins (OMPs) with heparin. Several bacterial strains showed differences in their profile of heparin-binding OMPs, but all exhibited low mol. wt (< 30 kDa) reactive proteins. Reactivity of whole bacterial cells with heparin was also observed by transmission electron microscopy. These results suggest that HSP are potential receptors for P. aeruginosa adherence to non-polarised and untight epithelial respiratory cells.
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PMID:Role of heparan sulphate proteoglycans as potential receptors for non-piliated Pseudomonas aeruginosa adherence to non-polarised airway epithelial cells. 1121 Dec 27