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)

We studied the interaction between glycosaminoglycans (GAGs) and fibronectin in the basement membrane of the epiblast in the chicken blastoderm using testicular-hyaluronidase digestion of GAGs either on fixed tissue sections or in vivo after microinjection of the enzyme preparation prior to immunostaining for fibronectin. In the choice of fixatives, special attention was paid to their preservation of GAGs. The controls included alcian-blue staining of serial sections to test the efficiency of the digestion, and incubations in the presence of protease inhibitors to abolish contaminating proteolytic activity in the commercial hyaluronidase preparations. The results indicate that fixation in solutions which preserve GAGs, i.e. ethanolic solutions or aqueous solutions containing cetylpyridinium chloride, allows the immunocytochemical demonstration of fibronectin in the basement membrane of the epiblast at the level of the endophyllic crescent, but masks this glycoprotein at the epithelial-mesenchymal interface. As shown by both approaches, this masking of immunoreactivity is reversible. Moreover, the in vivo clearance of GAGs before fixation shows that the masking at the epithelial-mesenchymal interface is not an experimental peculiarity due to the use of a particular technique, but is the consequence of an interaction between GAGs and fibronectin in that particular area of the basement membrane that is used by mesoblast cells as a substrate for migration. The observation that fibronectin may be masked by GAGs in ethanol-fixed tissue--a commonly used fixation method--may require the re-evaluation of some negative results mentioned in the literature.
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PMID:Masking of antigenic sites of fibronectin by glycosaminoglycans in ethanol-fixed embryonic tissue. 388 30

Microbial hyaluronidase (EC 4.2.2.1) was isolated from the culture fluid of Staphylococcus aureus 0-15 with purification by precipitation with 1 volume of ethyl alcohol, chromatography on DEAE cellulose and ultrafiltration through DA type membranes with the pore size of 0.65 micron ("Millipore") and PM-10 membranes ("Amicon"). The specific activity of the enzyme averaged to 2700 turbidimetric units or 32130 IU. 6585-fold purification of the enzyme was performed. The optimum action on hyaluronic acid was observed at pH 5.0-6.5. Hyaluronidase was inhibited by Fe3+, Fe2+ and Cu2+, activated by Ca2+ and stabilized by 0.15 M NaCl. It was detected that the enzyme had two molecular forms with the isoelectric points of 5.4 and 6.5 and the molecular weights of 55 000 and 24 0000 D respectively. The glycoprotein nature of the enzyme was shown. The immobilized form of hyaluronidase on activated polyglucin, a soluble biocompatible polymer was prepared. The form is characterized by higher thermostability.
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PMID:[Purification and properties of staphylococcal hyaluronidase]. 396 93

A highly purified commercial preparation of bovine testicular hyaluronidase (GL enzyme, Hyalosidase) was labelled with 125iodine without measurable loss of enzyme activity. The labelled preparation was administered intravenously into rats and the serum half-life of hyaluronidase was determined by measurement of both radioactivity and enzyme activity. The short half-life of the enzyme in plasma could not be accounted for by excretion in the urine and bile. Tissue distribution studies showed that the major site of uptake was the liver (59.7% of the recovered dpm). This rapid uptake by the liver could be reduced significantly by the pre-administration of yeast mannan or ovalbumin (a mannose-terminated glycoprotein). This suggests that the uptake of hyaluronidase by the liver is mediated by a mannose-specific receptor. Very little radioactivity was found in the heart (0.2% of the recovered dpm).
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PMID:The fate of intravenously administered highly purified bovine testicular hyaluronidase (Hyalosidase) in the rat. 400 38

Two collagen-poor, ultramicroscopic layers are described at the surface of canine articular cartilage. They are distinguished by staining with an electron-dense cationic dye, Cupromeronic Blue, in a critical electrolyte concentration technique and by digestion with testicular hyaluronidase. The superficial layer, approximately 50 nm thick, stained at low electrolyte concentrations but failed to stain in conditions specific for sulphated glycosaminoglycans. It was hyaluronidase-resistant and may be either glycoprotein or protein in nature. The deeper layer, 100-400 nm thick, stained positively at electrolyte concentrations specific for sulphated glycosaminoglycans but not in conditions specific for keratan sulphate. It was removed by hyaluronidase digestion. This layer probably represents a chondroitin sulphate-rich proteoglycan. These surface layers may be important in the lubrication of the articular surface and in the permeability and compression resistance of the superficial cartilage zone.
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PMID:Ultrastructural histochemistry of the surface lamina of normal articular cartilage. 401 50

The middle ear effusion specimens were obtained by myringotomy and aspiration from 4 children of 4-7 years old, who had been diagnosed as patients with secretory otitis media on the basis of conductive hearing loss and tympanogram. In cases 1 and 2, their ear fluids were macroscopically serous, while those of cases 3 and 4 were mucous. These ear fluids were digested with pronase and the digests were analyzed by cellulose acetate membrane electrophoresis with alcian blue and high-iron-diamine stainings. All samples were found to contain glycopeptides possibly derived from sulfated mucin-type glycoproteins with small amounts of glycosaminoglycans. The glycoconjugates from cases 3 and 4 were further examined after hyaluronidase and chondroitinase ABC treatments, followed by heparitinase digestion. The resultant glycopeptide fractions appeared to be electrophoretically homogeneous and their chemical compositions suggested that they were typical mucin-type glycopeptides. Furthermore, they contained sulfates. The data suggest that in secretory otitis media, one of the major components of middle ear effusions is sulfated mucin-type glycoprotein.
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PMID:Sulfated glycopeptides from middle ear effusions of secretory otitis media. 407 40

The radioautographic distribution of the label of galactose-H(3) was compared with that of glucose-H(3) in a series of secretory cells of the rat. Whereas the glucose label appeared in all mucous cells, the galactose label was incorporated only into certain mucous cells. Whenever either label was incorporated, however, it was located first in the Golgi region and later in the secretion product, mucus. Several lines of evidence, including extraction of glucose label with peracetic acid-beta glucuronidase, indicated that the material synthesized in the Golgi region was glycoprotein in nature. In chondrocytes, both the galactose and the glucose label appeared first in the Golgi region and later in cartilage matrix; extraction of glucose label with hyaluronidase indicated that much of it consisted of mucopolysaccharide. In all secretory cells, the extraction of glycogen by amylase had no effect on Golgi radioactivity. Such extraction did not eliminate the scattered cytoplasmic label also seen after glucose-H(3) injection, but completely eliminated that seen after galactose-H(3). Consequently, the galactose-H(3) label in the Golgi region stood out more clearly, and was detected in many cells: pancreas, liver, epididymis, and intestinal columnar cells. In the latter, label later appeared in the surface coat. Thus, radioautography after injection of galactose-H(3), as after glucose-H(3), indicates that synthesis of complex carbohydrates takes place in the Golgi region of many secretory cells.
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PMID:Radioautographic comparison of the uptake of galactose-H and glucose-H3 in the golgi region of various cells secreting glycoproteins or mucopolysaccharides. 422 8

Using affinity chromatography and enzyme-labelled immunological assays combined with affinity adsorption, we have obtained evidence for the binding of a brain glycoprotein to hyaluronic acid, and on this basis named it hyaluronectin. This binding was inhibited by hyaluronic acid and by the products of its hydrolysis by hyaluronidase from bovine testis, but was not inhibited by other glycosaminoglycans or by monosaccharides. Preparative affinity chromatography of brain acid-soluble proteins produced hyaluronectin in a good degree of purity. Contamination by albumin was less than 1% and the yield was as high as 80%.
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PMID:Characterization and purification from human brain of a hyaluronic acid-binding glycoprotein, hyaluronectin. 616 16

The role of osteoclasts or chondroclasts in degradation and synthesis of complex carbohydrates was investigated using the high iron diamine-thiocarbohydrazide-silver proteinate method (HID-TCH-SP) for sulfated glycoconjugates and the periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method for vicinal glycol-containing glycoconjugates. HID-TCH-SP stained the calcified cartilage matrix, the osteoclast ruffled border, vacuoles and heterophagosomes but not the Golgi apparatus and primary lysosomes. The size and number of HID-TCH-SP stain deposits progressively decreased from the calcified cartilage matrix to the ruffled border (p less than 0.001). Enzyme digestion with testicular hyaluronidase removed most HID-TCH-SP stain deposits averaging 13 nm. in diameter in the extracellular matrix, cytoplasmic vacuoles, and heterophagosomes of osteoclasts. Only sparse stain deposits averaging 8 nm. in diameter and presumed to be keratan sulfate remained in these sites after enzyme digestion. Osteoclast heterophagosomes contained the highest concentration of hyaluronidase-resistant material suggesting delayed degradation of keratan sulfate at this site. PA-TCH-SP strongly stained collagen fibrils in the calcified cartilage matrix. Reactive collagen fibrils were also observed in extracellular channels but only rarely were identifiable collagen fibrils observed in cytoplasmic vacuoles. A progressive decrease in the diameter of PA-TCH-SP reactive collagen fibrils was observed between the calcified cartilage matrix and the ruffled border region (p less than 0.001). PA-TCH-SP stained cisternae of rough endoplasmic reticulum, Golgi saccules, and primary lysosomes consistent with the synthesis and packaging of glycoprotein enzymes at these sites. These results indicate that the dissolution of sulfated glycoconjugates requires osteoclastic engulfment of degraded material and subsequent intracellular digestion, whereas the dissolution of collagen fibrils appears to be completed extracellularly.
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PMID:Extracellular and intracellular digestion of complex carbohydrates by osteoclasts. 617 27

A fetal antigen (FA) was isolated from spent culture medium of a melanoma (M14) cell line. Allogeneic serum samples from melanoma patients, previously characterized with respect to anti-FA activity, were used as the source of anti-FA antibody. The FA activity was partially purified by membrane ultrafiltration, gel filtration, and chloroform:methanol extraction. The partially purified FA was then used to develop an enzyme-linked immunosorbent assay (ELISA). By indirect ELISA both the IgG and IgM classes of anti-FA antibodies were detected in the sera of cancer patients and normal volunteers. The incidences of anti-FA antibodies in the sera of cancer patients and normal volunteers were not significantly different. As detected by competitive inhibition in ELISA, FA activity was widely distributed among melanoma, sarcoma, and carcinoma tumor tissues and cultured tumor cells, as well as among fetal brain, skin, and muscle tissues. FA activity was destroyed by treatment with beta-galactosidase and hyaluronidase, but it was not destroyed by proteolytic and lipolytic enzymes. The antigen bound to immobilized ricin, peanut, and soybean lectins. FA activity in material purified by ricin-affinity chromatography was associated with molecules in the 60,000- to 70,000-dalton region as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results suggest a glycoprotein nature for the FA isolated from the spent culture medium of melanoma (M14) cells; this FA apparently elicits formation of natural antibodies in the cancer patients and normal donors.
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PMID:Immunochemical characterization of fetal antigen isolated from spent medium of a human melanoma cell line. 619 35

Glycoproteins and proteoglycans synthesized by human keratinocytes in medium containing D-[1-14C]glucosamine were extracted and analysed by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. Extraction of the labelled keratinocytes with 0.5% Triton X-100 removed most of the glycoconjugates and left the cytoskeleton and nuclear residue adherent to the substratum. In addition to the cytoskeletal proteins, there was a relatively simple profile of glycoproteins and glycosaminoglycans associated with this adherent cytoskeleton. These consisted of eight glycoproteins in the mol.wt. range 99000-232000, five proteins in the keratin region (mol.wt. 42000-61000), hyaluronic acid and a sulphated glycosaminoglycan. Surface labelling of the keratinocytes with galactose oxidase (with or without neuraminidase)/KB3H4 revealed that many of the glycoproteins were exposed on the cell surface. The importance of the glycoproteins and proteoglycans in attaching the keratinocytes to the substratum was examined by studying their expression after incubation in medium containing tunicamycin and their degradation after digestion with trypsin and hyaluronidase. These studies, together with an examination of the glycoconjugates released by sequential extraction with 0.5% Triton X-100 followed by 0.2% sodium dodecyl sulphate, revealed that the glycoprotein of mol.wt. 232000 has an important role in mediating the attachment of keratinocytes to the substratum.
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PMID:Glycoproteins and glycosaminoglycans synthesized by human keratinocytes in culture. Their role in cell-substratum adhesion. 619 5


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