Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.6.4 (chondroitinase)
 2,039 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We examined immunocytochemically the type and distribution of glycosaminoglycans and proteoglycans (PG) in predentin and dentin demineralized with EDTA after aldehyde fixation of rat incisors using (a) four monoclonal antibodies (1-B-5,9-A-2,3-B-3, and 5-D-4) which recognize epitopes in unsulfated chondroitin (C0-S), chondroitin 4-sulfate (C4-S), chondroitin 6-sulfate (C6-S), and keratan sulfate (KS) associated with the PG, and (b) monoclonal (5-D-5) and polyclonal antibodies specific for the core protein of large and small dermatan sulfate (DS) PG. Light microscope immunoperoxidase staining after pre-treatment of tissue sections with chondroitinase ABC localized the majority of stainable PG (C4-S, KS, DSPG, C0-S, and C6-S) in predentin and, to a lesser extent (C4-S and small DSPG), in the dentin matrix. The former site demonstrated relatively homogeneous PG distribution, whereas the latter site revealed that strong staining of C4-S and small DSPG was confined mostly to dentinal tubules surrounding odontoblastic processes, with only weak staining in the rest of the dentin matrix. These results indicate that there is not only a definite difference between PG of predentin and dentin but also a selective decrease in the concentration or alteration of these macromolecules during dentinogenesis and mineralization.
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PMID:Immunohistochemical localization of glycosaminoglycans and proteoglycans in predentin and dentin of rat incisors. 168 34

We examined biochemically and immunocytochemically the type and distribution of mineral binding proteoglycans (PGs) in rat mid-shaft subperiosteal bone using three monoclonal antibodies (MAb 1-B-5, 9-A-2, and 3-B-3) which specifically recognize unsulfated chondroitin, chondroitin 4-sulfate (C4-S) and dermatan sulfate (DS), and chondroitin 6-sulfate. Bone proteins were extracted from fresh specimens with a three-step technique: 4 M guanidine HCl (GdnCl), aqueous EDTA without GdnCl (E-extract), followed by GdnCl. Western blot analysis of SDS-polyacrylamide gel electrophoresis revealed that E-extract after chondroitinase ABC digestion reacted strongly with MAb 9-A-2 but not with MAb 1-B-5 or 3-B-3. After adehyde fixation, ethanolic trimethylammonium EDTA was used as a demineralizing agent for light and electron immunocytochemistry. This provided good retention of water-soluble PGs in the specimens. After chondroitinase ABC pre-treatment of tissue sections, MAb 9-A-2 specifically stained C4-S and/or DS in the walls of osteocyte lacunae and bone canaliculi in the mineralized matrix as well as in the unmineralized matrix such as pre-bone, vascular canals, and pericellular matrix surrounding osteocytes; the remainder of the mineralized matrix lacked staining. These results indicate that mineral binding PGs contain C4-S and/or DS and are exclusively localized in the walls of the bone lacuna and canaliculus.
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PMID:Biochemical and immunocytochemical characterization of mineral binding proteoglycans in rat bone. 189 98

Proteoglycan molecules in keratoconus corneas were studied by immunohistochemical and electron microscopic histochemical methods. Compared with normal human control subjects, the staining intensity with monoclonal antibody 9-A-2 was enhanced in the stroma of scarred keratoconus corneas, whereas the intensity with antibody J-19 was reduced. The 9-A-2 experiment showed an increased immunoreactivity of dermatan sulfate proteoglycan epitopes, and the J-19 experiment indicated a decreased immunoreactivity of sulfated keratan sulfate epitopes. Uronic acid analyses were consistent with the 9-A-2 data. Electron microscopy performed after cuprolinic blue staining showed apparent accumulation of abnormally thick, chondroitinase ABC-sensitive, dermatan sulfate proteoglycan filaments in keratoconus corneas. Such filaments were especially prominent in scarred areas. In addition, Keratan sulfate proteoglycan filaments appeared to be less abundant than those found in normal control corneas. Similar alterations of both types of proteoglycan molecules were also seen and reported in scarred corneas. The similarity suggests that the proteoglycan abnormalities found in keratoconus corneas may be secondary, at least in part, to scarring.
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PMID:Proteoglycan molecules in keratoconus corneas. 203 6

A 68-year-old man and a 66-year-old woman had diffuse corneal stromal deposits that stained with alcian blue and colloidal iron but did not react with periodic acid-Schiff stain and lipid stains. Similar deposits were found within postmortem sclera in one case, but not in other ocular or extraocular tissues. The abnormal material was sensitive to testicular hyaluronidase and chondroitinase. The material reacted with monoclonal antibody 9-A-2 after digestion by chondroitinase AC in one case and ABC in both cases, which is consistent with the identification of the glycosaminoglycans chondroitin 4-sulfate and dermatan sulfate. Electron microscopic examination of the cornea in both cases disclosed granular material in vacuoles dispersed extracellularly and, rarely, in keratocytes. Results of blood and skin fibroblast enzyme assays for clinically relevant mucopolysaccharidoses and mucolipidoses were normal in both patients, and there were no somatic abnormalities suggesting a storage disease.
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PMID:Unusual mucopolysaccharide disorder with corneal and scleral involvement. 211 Apr 15

The proteoglycans associated with the mineralized matrix of bovine cementum have been studied biochemically and their distribution within this tissue localized immunohistochemically. Both hyaluronate and proteoglycans were fractionated by DEAE-Sephacel ion-exchange chromatography. The proteoglycans eluted in three separate peaks of which two contained alkali labile protein associated with glycosaminoglycans, and one appeared as free glycosaminoglycan chains. Analysis of the glycosaminoglycans identified chondroitin sulfate as the predominant species, although minor quantities of dermatan sulfate and heparan sulfate were also identified. Agarose-acrylamide gel electrophoresis and Sepharose CL-6B molecular sieve chromatography of the proteoglycans indicated them to be smaller in size with respect to periodontal ligament and gingival proteoglycans, but similar to bone and dentine proteoglycans. Amino acid analyses indicated subtle differences between cementum and bone proteoglycans. Using a monoclonal antibody (9-A-2) which recognizes the unsaturated disaccharide of chondroitinase ACII-digested glycosaminoglycans, chondroitin sulfate was identified in the pericellular environment within the lacunae housing the cementoblasts as well as in the extracellular matrix of cementum.
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PMID:Proteoglycans of bovine cementum: isolation and characterization. 211 81

Recent results show that type IX collagen isolated from chicken cartilage is associated with one or perhaps two chondroitin sulfate chains. To locate the chondroitin sulfate chain(s) along the type IX collagen molecule, rotary shadowing was performed in the presence of monoclonal antibodies which recognize stubs of chondroitin sulfate generated after chondroitinase ABC digestion. Monoclonal antibodies 9-A-2 and 2-B-6 which recognize stubs of chondroitin 4-sulfate were found to bind specifically to the NC3 domain of type IX collagen, and this binding was dependent on prior digestion of the preparation with chondroitinase ABC. Monoclonal antibody 1-B-5, which recognizes unsulfated stubs of chondroitin sulfate, did not show any specific binding to type IX collagen either with or without chondroitinase ABC digestion. As a control, monoclonal antibody 2C2 was used, which in previous work was shown to bind specifically to an epitope located close to or at the NC2 domain. Binding of this antibody to NC2 was unaffected by chondroitinase ABC digestion, and no specific binding of the antibody to the NC3 domain was detected either before or after chondroitinase ABC digestion.
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PMID:Use of monoclonal antibodies to locate the chondroitin sulfate chain(s) in type IX collagen. 309 5

The glycosaminoglycans that exist in rabbit bone marrow were analyzed chemically, and their in situ localization was studied immunohistochemically. Femoral bone marrow of 3-month-old rabbits was defatted with organic solvents. Glycosaminoglycans were prepared from the defatted tissue after its digestion with pronase, treatment with mild alkali, and then digestion with DNase-I. The tissue contained glycosaminoglycans equivalent to 195 mg of hexosamine per femur, which accounted for 27.3% of the total hexosamine in the tissue. Studies with hyaluronidase from Streptomyces hyalurolyticus and chondroitinase ABC showed that the glycosaminoglycans were composed of hyaluronic acid (16% of the total glycosaminoglycan) and chondroitin 6-sulfate (79%). The chondroitin 6-sulfate was separated on Bio-Gel A-0.5m gel into two molecular species with mol wt of greater than 12,000 (Kd greater than 0.2) and approximately 8,000 (Kd = 0.47). Bone marrow digested with chondroitinase ABC and then treated with three monoclonal antibodies 4/8/9-A-2, 5/6/3-B-3, and 5/6/1-B-5, which were specific for unsaturated 4-sulfated, 6-sulfated, and nonsulfated disaccharide structures, respectively, at the nonreducing end of chondroitin sulfate chains, reacted with only 5/6/3-B-3. This result indicated that the chondroitin sulfate, isomer in the bone marrow is chondroitin 6-sulfate, consistent with the biochemical results. The chondroitin 6-sulfate was localized mainly in the extracellular compartment and was considered to be involved in construction of the hemopoietic microenvironment in the bone marrow.
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PMID:Isolation, characterization, and localization of glycosaminoglycans in rabbit bone marrow. 311 61

Several monoclonal antibodies which recognize different antigenic determinants of chondroitin sulfate proteoglycan were used to study chondroitin sulfate proteoglycan biosynthesis in chicken chondrocyte cultures. The intracellular sites of synthesis and processing and extracellular deposition in matrix were localized by double immunofluorescence reactions. One rat monoclonal antibody, S103L , which recognizes an antigenic determinant of the core protein of the chicken cartilage chondroitin sulfate proteoglycan monomer, was used to identify both extracellular chondroitin sulfate proteoglycan and intracellular compartments containing chondroitin sulfate proteoglycan precursors. Intracellular staining with S103L was localized to perinuclear regions, and, in some chondrocytes, to a few other cytoplasmic vesicles as well. When chondrocytes were not fed for several days, intracellular chondroitin sulfate proteoglycan precursors were accumulated in larger compartments distributed throughout the cytoplasm. Polyclonal chondroitin sulfate proteoglycan antibodies displayed similar staining characteristics. In contrast, several of the monoclonal antibodies, including the rat monoclonals S11D and P100D , and the mouse monoclonals 1-B-5, 3-B-3 and 9-A-2, did not recognize native chondroitin sulfate proteoglycan, but reacted only with chondroitinase ABC-digested (and/or hyaluronidase-digested) chondroitin sulfate proteoglycan. These antibodies were particularly useful in the demonstration of the extracellular codistribution of chondroitin sulfate proteoglycan with either type II collagen or fibronectin. In other experiments, the monoclonal antibodies to chondroitin sulfate proteoglycan served to demonstrate that the perinuclear subset of intracellular compartments is uniquely involved in the addition of chondroitin sulfate oligosaccharides to the chondroitin sulfate proteoglycan core protein. Lastly, using the mouse monoclonal 5-D-4, which recognizes keratan sulfate determinants, the perinuclear region was identified as the site for keratan sulfate addition. Results suggest heterogeneity of keratan sulfate synthesis at the level of individual chondrocytes, even for cells apparently containing equivalent amounts of intracellular chondroitin sulfate proteoglycan.
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PMID:Immunofluorescence studies of chondroitin sulfate proteoglycan biosynthesis: the use of monoclonal antibodies. 620 57