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)

The biosynthesis of glycosaminoglycans (GAG) and glycopeptides was studied in rat kidney cortex, glomeruli, and isolated glomerular basement membranes (GBM). Rats were given four intraperitoneal injections of [(35)S]sulfate and [(3)H]glucosamine (over 10 hr) and sacrificed 14 hr after the last injection. Fractions of kidney glomeruli and purified GBM were prepared. The percent of the label incorporated into specific GAG or into glycopeptides was determined by selective degradative techniques in conjunction with gel filtration chromatography using the methods of Hart [Hart, G. W. (1976) J. Biol. Chem. 251, 6513-6521; Hart, G. W. (1978) Dev. Biol. 62, 78-98]. After digestion with Pronase and chromatography on Sephadex G-50, approximately 68% of the total (35)S radioactivity and 10-15% of the total (3)H radioactivity incorporated into cortex, glomeruli, or GBM was found in the GAG fraction, and the remainder ( approximately 32% of (35)S radioactivity and 85-90% of the (3)H radioactivity) was found in glycopeptide fractions. Treatment of GAG fractions isolated from the three sources (cortex, glomeruli, and GBM) with nitrous acid (which degrades heparan sulfates) indicated that the majority (85%, 65%, and 87%) of the (35)S radioactivity as well as the majority (60%, 50%, and 91%) of the (3)H radioactivity from all three sources was degraded by this treatment. When nitrous acid-resistant GAG from GBM were subjected to digestion with Streptomyces hyaluronidase (which degrades hyaluronic acid), approximately 6% of the (3)H-labeled material was sensitive to this treatment. The remaining (35)S- and (3)H-labeled GAG isolated from GBM were digested with chondroitinase ABC (which degrades chondroitin sulfates A and C and dermatan sulfate). Although the ratios of the types of GAG synthesized by all three sources were similar, in GBM the ratios of (35)S- to (3)H-labeled GAG and of (3)H-labeled glycopeptides to (3)H-labeled GAG were higher (2.5 times) than those found for glomeruli. The data demonstrate the synthesis of both sulfated and nonsulfated GAG by rat kidney cortex and glomeruli and their transport to and incorporation into the GBM. Heparan sulfate is the major GAG synthesized by glomeruli, but the glomeruli also synthesize smaller amounts of hyaluronic acid and chondroitin sulfates, which are in part incorporated into GBM. In addition, the renal cortex and the glomeruli synthesize glycopeptides, some of which are sulfated, and incorporate them into GBM.
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PMID:Sulfated and nonsulfated glycosaminoglycans and glycopeptides are synthesized by kidney in vivo and incorporated into glomerular basement membranes. 701 44

Extraction of rat glomerular basement membrane, purified by osmotic lysis and sequential detergent treatment, with 8 M urea containing protease inhibitors solubilizes protein that is devoid of hydroxyproline and hydroxylysine. This material represents 8-12% of total membrane protein, elutes mainly as two high molecular weight peaks on agarose gel filtration, and is associated with glycosaminoglycans. Isolated rat renal glomeruli incorporate [35S]sulfate into basement membrane from which this non-collagenous 35S-labeled fraction can be subsequently solubilized. The radioactivity incorporated into urea-soluble glomerular basement membrane eluted primarily with the higher molecular weight peak (Mr greater than 250 000). Cellulose acetate electrophoresis after pronase digestion of the urea-soluble fraction revealed glycosaminoglycan that was resistant to digestion with Streptomyces hyaluronidase and chondroitinase ABC, sensitive to nitrous acid treatment, and contained [35S]sulfate. The findings indicate that one of the non-collagenous components of glomerular basement membrane is a proteoglycan containing heparan sulfate.
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PMID:Non-collagen protein and proteoglycan in renal glomerular basement membrane. 731 55

The subcellular route of incorporation of complex carbohydrates into rabbit heterophil primary granules and their subsequent intragranular distribution during granule maturation were studied with ultrastructural, cytochemical, and radioautographic methods. High iron diamine (HID) staining of sulfated glycoconjugates in primary granules was partially diminished after treatment with chondroitinase ABC or after removal of N-sulfate groups with nitrous acid, but was not altered by exposure to hyaluronidase, trypsin, or HCl. Subsequent thiocarbohydrazide-silver proteinate (TCH-SP) straining of thin sections increased the density of the HID reaction product. Golgi-derived spherules and very immature morular granules stained weakly with HID-TCH-SP and labeled intensely after a 10 min incubation with 35SO4. After a 60 min 35SO4 pulse and a 60 min chase, an increase in radiolabeling was observed in granules with HID stained, fused morular material, and some labeling was present in more mature rim stained granules. Fully mature granules lacked HID or HID-TCH-SP staining, but contained most of the 35SO4 labels after a 60 min pulse and 18 hr chase in vitro. Periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) staining of unosmicated thin sections localized vicinal glycol-containing complex carbohydrates in Golgi-associated small vesicles. These vesicles lacked HID-TCH-SP staining and apparently contained neutral glycoprotein. They frequently bordered, in a rosette arrangement, the immature morular granules, but not the more mature primary granules. The PA-TCH-SP method localized complex carbohydrates in the rim of granules precursors and enclosed a spherule or morula, but failed to stain the sulfate-containing material in the morulas or spherules. PA-TCH-SP reactivity was diffusely distributed in moderately mature granules and was decreased in fully mature granules. These results indicate that heterophil primary granule contain several complex carbohydrates including O-sulfated and N-sulfated glycosaminoglycans, as well as vicinal glycol-containing glycoproteins. These complex carbohydrates are transported to immature primary granules by different Golgi-derived organelles. The complex carbohydrates are subsequently distributed differently within primary granules and become masked to staining as the granule matures.
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PMID:Ultrastructural cytochemistry and radioautography of complex carbohydrates in heterophil granulocytes from rabbit bone marrow. 741 99

The localization and the nature of glycosaminoglycans (GAGs) in the synovial membrane in 30 patients with rheumatoid arthritis (RA) involving 40 knees were studied by newly developed histochemical methods. To detect the acidic glycoconjugates, sensitized diamine procedures were employed based upon high and low iron diamine stainings. To identify the various molecular species of the GAGs, enzyme (chondroitinase ABC and B, testicular hyaluronidase and keratanase) digestion and chemical modification (nitrous acid treatment) procedures were performed prior to the diamine stainings. The sensitized diamine methods could clearly stain the acidic glycoconjugates contained in the synovial tissue components in shades of brown to black, and could detect the precise distribution patterns of the GAGs. The results obtained in the present study confirmed that the tissue in RA synovial membranes contained various amounts of each GAG molecular species such as dermatan sulfate, chondroitin sulfate A/C, hyaluronic acid and heparan sulfate. Furthermore, the distribution patterns of dermatan and chondroitin sulfates in the diseased synovial tissues were pathophysiologically interesting; in the inflammatory areas, the molecular species of GAGs was primarily dermatan sulfate, whereas in the fibrotic areas, it was mainly chondroitin sulfate A/C. Such results appear to be useful for pathophysiological studies on the synovial tissues of RA.
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PMID:[A histochemical study on the acidic glycoconjugates of the synovial membrane in rheumatoid arthritis]. 818 1

The molecular weight distribution of pMP-derived glycosaminoglycans (GAG), i.e. non-sulfated GAG, chondroitin sulfate (CS), and heparin sulfate (HS)-like material was determined. The peritoneal macrophages (pMP) were harvested from rats normal or stimulated by i.p. injection of thioglycolate, carrageenan or BCG, and maintained in culture. The GAG of cell layer and medium were isolated separately after labeling with 35S-sulfate and 3H-acetate. Treatment with nitrous acid served to remove HS-like material. Labeling with 3H-acetate served to detect synthesis of the high m. w. hyaluronic acid (HA). Gel chromatic separation was done using Sephadex G-200 columns. The maximal size of 35S-labeled GAG, especially HS (36 kDa), was reduced in cultural medium and cell layer after stimulation in vivo. Reduction was most pronounced after application of carrageenan followed by thioglycolate and BCG/LPS stimulation. The extracellular GAG of BCG-stimulated pMP were smallest, probably due to degradation. Heparan sulfate-like material made up a larger proportion in monolayer and medium, comprising the total m.w. range up to 36 kDa. The GAG sensitive to nitrous acid were maximal in cultures of carrageenan-stimulated pMP and minimal in those of thioglycolate-stimulated pMP. This type of HS was sensitive to hyaluronidase, too. Any synthesis of high molecular hyaluronic acid was not found in normal or stimulated rat pMP. Therefore MP-associated HA must be adsorbed from other sources or synthesized by early forms of macrophages.
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PMID:The glycosaminoglycans in cultures of stimulated rat peritoneal macrophages. 2. Gel chromatographic studies and the behaviour of heparan sulfate. 832 74

HPLC-purified glycosaminoglycans (hpGAG) prepared from extracts of non-luteal mouse (JcL:ICR strain) ovaries were assayed for neovascularization by implanting Elvax films, containing test samples, on the lateral wall of the sheath of m. rectus abdominis in adult female mice of the same strain. Neovascularization occurred in a dose-dependent manner, and was characterized by capillary outgrowth extending into the tissue surrounding the implant. The single major peak of purified GAG on a column of TSK gel DEAE got out of order after treatment with streptococcal hyaluronidase or nitrous acid. The activity of this fraction was also greatly reduced when treated with streptococcal hyaluronidase or nitrous acid. When hpGAG was embedded in the implant with 17 alpha-hydroxyprogesterone at a dose of 20 micrograms/film, neovascularization induced by means of hpGAGs was suppressed. Progesterone at a dose of 50 micrograms/film did not suppress the neovascularization induced by ovarian hpGAG. These findings suggest that 17 alpha-hydroxyprogesterone suppresses the angiogenic activity of hyaluronic acid-like hpGAG in the ovary.
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PMID:17 alpha-Hydroxyprogesterone suppresses neovascularization induced by HPLC-purified ovarian hyaluronic acid-like glycosaminoglycan in mice. 857 92

The structural organization of integral and associated components of the ciliary zonule is still not fully understood. The present study is to localize and characterize the proteoglycans associated with the ciliary zonule of the rat eye by Cuprolinic blue (CB) staining and immunocytochemistry. After CB staining, the proteoglycans appeared as electron dense elongated rodlets and were localized with the zonular fibers. They were seen lying on the periphery of the zonular fibers or along the length of the individual fibrils. Most of the CB rodlets had a size of 60-170 nm long (average 130 nm) and 25 nm wide. Smaller CB rodlets measuring 25-60 nm long (average 45 nm) and 12 nm wide were sometimes found associated with the individual zonular fibrils. The CB rodlets were removed after chondroitinase ABC or chondroitinase AC treatment, but were resistant to heparitinase, nitrous acid, keratanase or Streptomyces hyaluronidase digestions. The ciliary zonule was also immunostained with three monoclonal antibodies: 2-B-6 specific for chondroitin 4-sulfate, 3-B-3 for chondroitin 6-sulfate and 1-B-5 for unsulfated chondroitin, using indirect immunoperoxidase or immuno-colloidal gold methods. The zonular fibers were immunoperoxidase stained and immunogold labeled by 2-B-6, but were not reactive to 3-B-3 and 1-B-5. The results demonstrate that chondroitin sulfate proteoglycan is associated with the ciliary zonule of the rat eye.
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PMID:Proteoglycans associated with the ciliary zonule of the rat eye: a histochemical and immunocytochemical study. 857 87

Histochemical studies were carried out in the mouse on the changes and distribution patterns of glycosaminoglycan molecular species during the separation of the lens vesicle. To obtain embryos, pregnant mice of the Jcl:ICR strain were sacrificed on day 10.5 or 11 of pregnancy. Serial frontal sections were strained with hematoxylin-eosin and sensitized high iron diamine. To identify glycosaminoglycan molecular species in tissues, an enzyme digestion (double digestion with chondroitinase B and testicular hyaluronidase) and a chemical modification (nitrous acid treatment) were performed in combination with the sensitized high iron diamine method. Before separation of the lens vesicle, chondroitin sulfate A/C and B were found in the basement membrane of the future corneal epithelium and intercellular matrices between the future corneal epithelium and lens vesicle, and chondroitin sulfate A/C was identified in the lens capsule. After separation of the lens vesicle, heparan sulfate emerged in the basement membrane of the future corneal epithelium and intercellular matrices between the future corneal epithelium and lens vesicle. These results indicate that the changes and distribution pattern of glycosaminoglycan molecular species play an important role during the separation of the lens vesicle.
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PMID:Histochemical studies of the separation of the lens vesicle in the mouse. 892 40

The glycosaminoglycans of sciatic nerves recovering from crush-injury were studied in adult guinea pigs and compared with those of non-injured mature neural tissues. The glycosaminoglycans were recovered from the 1,900 g supernatant and pellet of the tissue homogenates and assayed for hexuronate contents and susceptibilities to hyaluronidase, chondroitinase ABC, and nitrous acid. In the normal brain and central nerve tracts, the glycosaminoglycans were distributed both in the supernatant and pellet fractions; the brain showed a predominance of chondroitin sulphates but the tracts showed a predominance of heparan sulphates. Twice as much glycosaminoglycans were found in normal sciatic nerves, only in the pellet fraction and with heparan sulphate predominant. In the 2 weeks post-crush, progressive increase in hexuronate was observed, due mainly to additional chondroitin sulphate forms in the supernatant; the pellet fraction in the same period was however similar to the untreated controls in relative abundance of glycosaminoglycan classes and hexuronate content. At 4 weeks post-crush, although the total hexuronate returned to the control level, a significant proportion of glycosaminoglycans remained in the supernatant fraction. Evidence is thus provided for the need to modulate the glycosaminoglycan expression pattern in adult neural tissue to allow post-traumatic tissue remodelling and axonal regrowth.
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PMID:Changes in glycosaminoglycans during regeneration of post-crush sciatic nerves of adult guinea pigs. 895 Jul 6

We studied histologically the changes and distribution patterns of glycosaminoglycan molecular species during the separation of the lens vesicle in the mouse. Embryos were obtained by sacrificing pregnant mice of the Jcl: ICR strain on day 10.5 and 11 of pregnancy. Serial frontal sections were stained with hematoxylin-eosin and a sensitized high iron diamine method. To identify glycosaminoglycan molecular species in tissues, enzyme digestion (double digestions with chondroitinase B and testicular hyaluronidase) and chemical modification (nitrous acid treatment) were performed in combination with the sensitized high iron diamine method. Before separation of the lens vesicle, the glycosaminoglycan molecular species, identified in the basement membrane of the presumed corneal epithelium and intercellular matrices between the presumed corneal epithelium and lens vesicle, were chondroitin sulfate A/C and B, and those in the lens capsule were chondroitin sulfate A/C. After separation of the lens vesicle, heparan sulfate emerged in the basement membrane of the presumed corneal epithelium and intercellular matrices between the presumed corneal epithelium and lens vesicle. These results are thus taken to indicate that the changes and distribution patterns of glycosaminoglycan molecular species play an important role during separation of the lens vesicle.
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PMID:[Histochemical studies on the separation of the lens vesicle]. 902 6


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