Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.2.1.36 (hyaluronidase)
4,606 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A morphologically detectable cell coat, composed of glycoprotein, glycolipid, and glycosaminoglycan, is present on the external surface of most vertebrate cells. We have investigated the composition and organization of glycosaminoglycans in the cell coat of cultured human embryo fibroblasts by labeling cells with 3H-glucosamine and Na235SO4 and subsequently treating cultures with specific enzymes. Components released were identified by chromatography and specific enzymatic digestion. In situ incubation with leech hyaluronidase (4 microgram/ml) removed only hyaluronic acid from the cell surface whereas testicular hyaluronidase (0.5 mg/ml) removed both hyaluronic acid and chondroitin sulfate. Trypsin (0.1 mg/ml) released a large mass of glycopeptides in addition to hyaluronic acid, chondroitin sulfate, and heparan sulfate. The affinity of the cell coat for the cationic dye, ruthenium red, was reduced by leech hyaluronidase treatment. Sequential enzyme digestions of the cell surface showed that hyaluronic acid could be removed without the concomitant or subsequent release of sulfated glycosaminoglycans, suggesting that the hyaluronic acid is not a structural backbone for glycosaminoglycan complexes of the external cell surface.
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PMID:Cell surface glycosaminoglycans: identification and organization in cultured human embryo fibroblasts. 90 84

Cellular glycosaminoglycans were isolated from lymphocytes from patients with cystic fibrosis and controls. The isolated glycosaminoglycans were fractionated by cellulose acetate electrophoresis, analyzed for glucosamine and galactosamine content, and subjected to hydrolysis with bovine testicular hyaluronidase. The total glycosaminoglycan content, the per cent glucosamine and galactosamine, and the distribution of cellular glycosaminoglycans in circulating lymphocytes in cystic fibrosis were no different from controls.
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PMID:Cellular glycosaminoglycans in lymphocytes from patients with cystic fibrosis. 100 Aug 40

The enzymes from the venom of Heterometrus scaber, the indole compounds present and the toxic protein of the venom have been studied. The venom contains acid phosphatase, ribonuclease, 5'-nucleotidase, hyaluronidase, acetylcholine esterase and phospholipase. A. The indole compounds present in the venom have been identified as 5-hydroxytryptophan, tryptophan, serotonin and tryptamine, along with two unidentified indole compounds. The venom produces hyperglycaemia in sublethal doses and this has been found to be due to increased adrenaline secretion. The toxic protein of the venom has been obtained in a pure form by (NH4)2SO4 fractionation, followed by fractional precipitation with acetone and chromatography over DEAE-Sephadex. The toxic fraction has been found to be homogeneous on acrylamide gel electrophoresis. It is a glycoprotein (molecular weight 15 000) containing 1.74% glucosamine, 0.87% galactosamine, 0.313% sialic acid, 3.25% fucose and 0.45% of an unidentified neutral sugar. It did not show any enzyme activities, haemolytic activity or inhibition of succinate dehydrogenase activity but it produced hyperglycaemia in sublethal doses. The toxic level (intravenous administration in rats) was found to be 0.72 mg/kg body weight.
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PMID:Investigations on the venom of the South Indian scorpion Heterometrus scaber. 111 82

A new biomaterial containing covalently bound hyaluronidase was prepared. An application of this enzyme membrane is to improve the performance of an implantable fuel cell. Hyaluronic acid is a contributor to the viscosity of tissue fluids but can be a potential fuel source because of its sugar content. The incorporation of immobilized hyaluronidase would not only contribute to a more available fuel supply by splitting hyaluronic acid but, perhaps more importantly, enhance the rate of mass transport of fuel, O2, and reaction products by reducing the viscosity near the electrode membranes. Hyaluronidase was bound to Sepharose gel and its thermoplastic membrane after activation by cyanogen bromide. Fourteen and 22% of the activities were recovered from the gel and membrane, respectively. The activity of the bound enzyme was stable for six months at 0 degrees C. The addition of hyaluronic acid, 1 mg/ml, to a typical implantable type bioautofuel cell in vitro increased external solution viscosity from 1.1 to 2.5-2.8 cP and reduced voltage output under 10 komega by 60% in 3 hr. When the hyaluronidase bound membrane was placed at the anode, viscosity of the glucose-hyaluronic acid solution was lowered to 1.8 cP and the cell output increased to the original level of a glucose-fueled cell in 3 hr. Glucosamine-equivalent released from hyaluronic acid at the electrode was 3.1 mg after 22.5 hr. This represents 90% of the theoretical consumption. Restoration of the cell output was probably a combination of the enhanced transport of fuel, O2 and products, and/or appearance of a new fuel, glucosamine-equivalent.
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PMID:Hyaluronidase-bound membrane as a biomaterial for implantable fuel cells. 125 16

Proteoglycans (PGs) from human burn hypertrophic scar of a patient with Ehlers-Danlos syndrome were extracted with 4M guanidinium chloride and purified by DEAE-cellulose chromatography. Differential ethanol precipitation of the PG fraction obtained after ion-exchange chromatography yielded two low mol.-wt. PGs, on rich in glucuronic acid (PGGLCA; Mr 66 kDa) and the other rich in iduronic acid (PGIDOA; Mr 48 kDa). In PGGLCA, 84% of the glycosaminoglycan chains are composed of GlcA----GalNAc(SO4) units, whereas in PGIDOA, the chains contain 95% IdoA----GalNAc(SO4) disaccharide units. Upon treatment with testicular hyaluronidase, the PGs gave different-sized oligosaccharides. Chondroitinase ABC digestion of PGGLCA or PGIDOA gave a single protein core (Mr approximately 20 kDa). The presence of glucosamine and sialic acid in PGGLCA and PGIDOA suggests that both contain N-linked oligosaccharides.
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PMID:Proteoglycans in human burn hypertrophic scar from a patient with Ehlers-Danlos syndrome. 159 19

A high molecular weight secretion product of cell lines established from duct cell-derived human pancreatic adenocarcinomas was investigated in this study. After metabolic labeling and molecular sieve chromatography of culture medium, a product appeared in the void volume of a Superose 6 column that could be labeled with [3H]glucosamine, but not with [35S]sulfate. After further purification by anion exchange chromatography it was analyzed and demonstrated to be hyaluronan (HA). CsCl density gradient centrifugation revealed a density of 1.45 g/cm3 in a 4 M guanidinium hydrochloride solution. [3H]Glucosamine-labeled material could be degraded by digestion with hyaluronidase from two sources, but not with heparitinase I or chondroitinase AC. Sugar analysis revealed glucuronic acid and glucosamine at a molar ratio of 1:1. When the amount of HA synthesized by different pancreatic adenocarcinoma cell lines was compared, the values of the cell lines PaTu 8902 and PaTu II were about five- to tenfold higher than those of the lines PaTu 8988s, PaTu 8988t or HPAF, but an order of magnitude lower than in murine 3T3 fibroblasts. HA synthesis per cell decreased with increasing cell density. In serum-free cultures of cell lines with high HA synthesis it was 3 to 5 times higher compared to cultures that were supplemented with serum. We conclude that pancreatic adenocarcinoma cells secrete hyaluronan and thus contribute to the extracellular matrix of the tumor tissue. In pancreatic carcinoma cells, regulation of HA biosynthesis seems not to be positively correlated to proliferation as has been demonstrated for fibroblasts.
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PMID:Hyaluronan is a secretory product of human pancreatic adenocarcinoma cells. 164 63

The synthesis and secretion of mucin-like high-molecular glycoprotein was studied in 2 human colon cancer cell lines that spontaneously differentiate in culture (Caco-2 and T84) and in 2 cell lines that do not spontaneously differentiate (LS174T and HT29). Mucin, quantitated by 3H-glucosamine labelling and chromatography on Sepharose CL-4B was found to be produced by all 4 cell lines. The mucinous nature of the labelled high-molecular glycoprotein was verified by enzymatic degradation treatments (heparinase, hyaluronidase, chondroitinase ABC, and N-glycanase), alkaline-borohydride treatment, inhibition of labelling by the glycosylation inhibitor benzyl-alpha-GalNAc, and by CsCl-density-gradient centrifugation. In all 4 cell lines, an inverse correlation of mucin synthesis with cell density was demonstrated. In Caco-2 cells, the spontaneous post-confluent enterocytic differentiation with increased brush-border enzyme expression was associated with a decrease in mucin synthesis and in the activities of polypeptidyl GalNAc transferase and beta 1,3-galactosyltransferase activity. Using cDNA probes for 2 distinct human intestinal mucins (MUC2 and MUC3), we found that all 4 colon cancer cell lines expressed mucin message, but the types of mucin mRNA expressed differed. These data indicate that mucin-like glycoproteins can be synthesized by cell lines derived from non-mucinous colon cancer, whether or not they undergo spontaneous differentiation in culture. These cell lines may serve as in vitro models for studying apomucin heterogeneity and control of mucin gene expression.
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PMID:Mucin synthesis and secretion in relation to spontaneous differentiation of colon cancer cells in vitro. 172 5

The membranous labyrinth of the inner ear, with its three semicircular canals, originates from a simple spheroidal otic vesicle. The process is easily observed in Xenopus. The vesicle develops three dorsal outpocketings; from the two opposite faces of each outpocketing pillars of tissue are protruded into the lumen; and these paired 'axial protrusions' eventually meet and fuse, to form a column of tissue spanning the lumen of the outpocketing like the hub of a wheel, with a tube of epithelium forming the semicircular canal around the periphery. Each axial protrusion consists of epithelium encasing a core of largely cell-free extracellular matrix that stains strongly with alcian blue. In sections, at least 60% of the stainable material is removed by treatment with Streptomyces hyaluronidase. When Streptomyces hyaluronidase is microinjected into the core of a protrusion in vivo, the protrusion collapses and the corresponding semicircular canal fails to form. Hyaluronan (hyaluronic acid) in the core of the protrusion therefore seems to be essential in driving the extension of the protrusion. Autoradiography with tritiated glucosamine indicates that the hyaluronan-rich matrix is synthesised by the epithelium covering the tip of the protrusion; the basal lamina here appears to be discontinuous. These findings indicate that the epithelium of the axial protrusion propels itself into the lumen of the otocyst by localised synthesis of hyaluronan. Hyaluronan may be used in a similar way in the development of other organs, such as the heart and the secondary palate.
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PMID:Hyaluronan as a propellant for epithelial movement: the development of semicircular canals in the inner ear of Xenopus. 179 22

The chain length of [3H]hyaluronic acid synthesized by cultivating human skin fibroblasts in the presence of [3H]glucosamine was investigated. [3H]Hyaluronic acid obtained from the matrix fraction was excluded from a Sepharose CL-2B column irrespective of the incubation period, whereas that from the medium was depolymerized into a constant chain length (Mr = 40,000). The reducing and non-reducing terminals of the depolymerized hyaluronic acid were N-acetylglucosamine and glucuronic acid, respectively. Prolonged incubation produced no oligosaccharides as shown by examination of hyaluronidase digests, suggesting the presence of a novel endo-beta-N-acetylglucosaminidase in cultured human skin fibroblasts.
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PMID:Extracellular depolymerization of hyaluronic acid in cultured human skin fibroblasts. 222 83

Assay conditions for determining hyaluronic acid levels in cultured cells have been examined. In cultures labeled with [3H]glucosamine, hyaluronic acid is measured by digestion with a highly specific hyaluronidase from Streptomyces hyaluronlyticus. Products obtained in the presence and absence of preliminary enzyme digestion are precipitated with cetylpyridinium chloride. The precipitation step has been optimized for ion concentration, glycosaminoglycan carrier and for cetylpyridinium chloride levels. Chondroitin sulfate is an effective carrier in the precipitation of radiolabeled product, while unlabeled hyaluronic acid is not. Addition of sulfate to the mixture yields a flocculent precipitate that facilitates subsequent steps of the determination. Optimizing these steps in hyaluronic acid determination can generate two- to three-fold increases in apparent levels of deposition in cultured cells.
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PMID:Hyaluronic acid determinations: optimizing assay parameters. 237 19


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