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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The biosynthesis and composition of glycosaminoglycans (GAG) in the endothelium-covered neointima, formed in response to de-endothelialization of the rabbit aorta by a balloon catheter, was examined. The [14C]glucosamine incorporation into GAG during an in vitro incubation with intimal-medial tissue was monitored periodically up to 24 hr. The GAG were isolated after an exhaustive proteolytic digestion with pronase and protease followed by ethanolic precipitation at 4 degrees C. Electrophoretic migration on cellulose acetate paper was compared for identification. The distribution of GAG was determined after a selective enzymatic digestion of isolated GAG using specific enzymes. Heparan sulfates were estimated after nitrous acid treatment. The concentration of GAG was measured spectrophotometrically by forming colored complexes with Alcian blue dye. In addition, the specific activity (dpm/microgram GAG) and the rate of GAG synthesis (ng/mg dry defatted tissue/day) were determined. The results indicate that the rate of GAG synthesis by de-endothelialized neointima (DEA) was twice that of intact aorta (control). In the re-endothelialized neointima (REA), the GAG synthetic rate was three times more than in control. However, the release of GAG into medium from REA accounts for only 25% of the GAG synthesized by this tissue type, and the release from DEA accounts for 60% of the synthesized GAG. Similarly, a threefold increase in the GAG concentration in REA compared to control was found. The relative distribution as chondroitin-6-sulfate (C6S), chondroitin-4-sulfate (C4S), dermatan sulfate (DS), heparan sulfate (HS) and hyaluronic acid (HA) was markedly altered in the injured neointima. There was an increase in chondroitin sulfates (CS) and DS concomitant with a decrease in HS. It is concluded that injury to aortic endothelium induces stimulation of GAG synthesis in the arterial wall. Furthermore, the greater release of GAG from DEA, compared to control and REA, suggests that endothelium may function as a "reverse" barrier in the neointima covered by regenerated endothelium.
Exp Mol Pathol 1985 Jun
PMID:Glycosaminoglycan composition and biosynthesis in the endothelium-covered neointima of de-endothelialized rabbit aorta. 392 84

Glycosaminoglycans (GAG) are believed to be important in the pathogenesis of atherosclerosis. We have previously demonstrated that areas of injured aorta that have been re-endothelialized accumulate increased amounts of lipid and GAG when compared to areas remaining de-endothelialized. We have now examined the net incorporation of labeled precursors into the individual GAG present in both re-endothelialized and de-endothelialized areas of rabbit aorta. Aortic tissue was examined at 2-3 and 10-14 weeks after a denuding injury by incubating tissue minces with [3H]glucosamine and sodium [35S]sulfate for 24 hr. Following incubation, the aortic GAG were isolated and assayed for uronic acid concentration and radioactivity. Results indicate that the total GAG concentration was significantly greater (P less than 0.001) in the re-endothelialized (9.46 +/- 0.29 micrograms/mg lipid-free dry residues (LFDR), mean +/- SE) as compared to de-endothelialized (7.89 +/- 0.43 micrograms/mg LFDR) areas. The concentration in uninjured aorta was 9.01 +/- 0.69. The difference between the injured tissues was attributable to increased concentrations of sulfated GAG. Hyaluronic acid and chondroitin sulfate were the most metabolically active of the GAG in either uninjured or injured aorta, together accounting for over 75% of the 3H label. The 3H specific radioactivities of the four GAG in the short-term, re-endothelialized subgroup were all increased nearly twice that found in uninjured and de-endothelialized tissues. With the exception of heparan sulfate, no significant differences were noted in the 3H specific radioactivities between the re-endothelialized and de-endothelialized areas in the long-term subgroup. These results indicate that, relative to adjacent areas of de-endothelialization, GAG preferentially accumulate in re-endothelialized areas even as early as 2-3 weeks following a denuding injury. Overall, metabolic data suggest that increased synthesis is responsible for this effect, although the net contribution of degradative processes cannot be overlooked since GAG turnover was not specifically examined. Thus, it is possible that regenerated endothelium may modify the GAG metabolism of the arterial wall following arterial injury.
Exp Mol Pathol 1985 Jun
PMID:Altered glycosaminoglycan metabolism in injured arterial wall. 399 53

We recently demonstrated that PC12 cells cultured on extracellular matrix (ECM) exhibit a flattened morphology, release more dopamine and contain less intracellular dopamine than cells which have a rounded shape on plastic culture ware. To further explore the role of PC12 cell shape in dopamine processing, we characterized the interaction of ECM and various agents on cell shape and dopamine release and content. In addition, the constituents of the ECM and the corneal endothelial cells which produce the ECM were presented in soluble form to PC12 cells on plastic. The soluble polysaccharides heparin, dextran and dextran sulfate, caused a dose-related increase in rounded (refractile) cells on ECM, a dose-related decrease in dopamine release and an increase in dopamine content relative to flattened cells on ECM. Chondroitin sulfate and hyaluronic acid did not prevent cell spreading on ECM nor affect dopamine processing. These experiments demonstrate that certain polysaccharides block cell spreading on ECM and that round cells on ECM secrete and store dopamine in a fashion similar to round cells on plastic. Colchicine, cytochalasin B and cycloheximide appear to affect dopamine synthesis and thus could not distinguish the role of cell shape in transmitter release. Treatment of PC12 cells on plastic with collagens I, II, III, IV, fibronectin, fibroblastic growth factor (FGF), or solubilized ECM had little effect on dopamine release or content. Endothelial cell conditioned medium and endothelial cell lysate increased dopamine release, but also increased dopamine content which differs from the effect of ECM. In summary, these experiments suggest that extracellular matrix alters cell shape and that the physical arrangement of these cells can determine dopamine secretion and storage.
Mol Cell Endocrinol 1985 Aug
PMID:Further characterization of substratum influence on PC12 cell shape and dopamine processing. 402 3

The in vitro interaction of hyaluronic acid (HA) with complement (C) classical-pathway activity has been investigated. It was found that native HA, even at a high concn (greater than 3 mg/ml), has a relatively weak anticomplementary activity. However, we report here that native HA can be reversibly altered by heat treatment such that C-inhibitory properties are manifested. We have determined in this study that a potent C-inhibitory activity can be obtained if HA solutions are thermally treated (100 degrees C), and stabilized by prompt freezing with prompt thawing just prior to the interaction with human serum complement. Several investigators have proposed that the intermolecular-associated strands of HA undergo a reversible decoupling upon thermal treatment and this decoupled state of HA can be semi-stabilized by quickly cooling the sample. This heat-treated HA strongly inhibits C1 as well as classical-pathway-mediated C3 conversion. However, if heat-treated HA samples are not stabilized but, rather, slowly cooled after heating or if heated HA samples are snapfrozen and then slowly thawed, the anticomplementary activity is gradually lost. Interestingly, the activity for this same sample can be regenerated by retreatment of the same sample with heat followed by low-temp stabilization, indicating the reversibility of the physical state of HA responsible for the anticomplementary effect. Since no detectable molecular degradation of thermally-treated HA was found, it was assumed that a heat-induced physical transition of HA (decoupled state) was responsible for the C-inhibitory effect.
Mol Immunol 1985 Apr
PMID:Hyaluronic acid-complement interactions--I. Reversible heat-induced anticomplementary activity. 403 63

Native hyaluronic acid (HA) is reported to be a weak anticomplementary agent. However, the normal buffer systems used for complement tests incorporate gelatin, Ca2+ and Mg2+, which may bind to HA, influence its conformation and interfere with its anticomplementary reactions with complement components such as Cl. In this study, metal ions (Ca2+ and Mg2+), gelatin and fibronectin appeared to react with native HA preparations and block their anticomplementary effects on Cl. In previous studies, we obtained evidence for a relationship between reversible heat-induced HA conformational changes and a subsequent reversible increase in anticomplementary activity. The anticomplementary activity of heat-treated HA preparations was also reduced by gelatin.
Mol Immunol 1985 Aug
PMID:Hyaluronic acid-complement interactions--II. Role of divalent cations and gelatin. 404 42

Bovine cumulus-oocyte complexes from small (1-5 mm) follicles were cultured for 24 h in 0.25 ml minimum essential medium supplemented with 10% fetal bovine serum and 20 microCi [3H]glucosamine. Treatment groups consisted of supplementing the culture medium with no hormone (control), 0.5 IU/ml follicle-stimulating hormone (FSH) or 10 mM 8-Br-adenosine cyclic monophosphate (cAMP). After culture, the complexes were fixed for light and scanning electron microscopy. Electron photomicrographs revealed that complexes induced to expand with FSH or cAMP contained a copious glycosaminoglycan (GAG) matrix extending between and around the cumulus cells. Control complexes did not exhibit expansion or an extracellular matrix. The radiolabeled GAG material was isolated for chemical identification. Chemical analyses included: (1) electrophoresis of GAG material, (2) electrophoresis of GAG material after enzyme or nitrous acid treatment, (3) thin-layer chromatography of GAG hydrolysates. The results from electrophoresis showed that the radiolabeled GAG co- migrated with hyaluronic acid. The GAG material was resistant to chondroitinase ABC and nitrous acid degradation but was digested by hyaluronidase. Complexes treated with FSH and cAMP incorporated higher (P less than 0.1 and P less than 0.025 respectively) amounts of [3H]glucosamine into hyaluronic acid than control cultures. Thin-layer chromatography identified the primary amino sugar of the GAG to be glucosamine. These data collectively showed that the radioactive GAG produced by bovine cumulus-oocyte complexes was hyaluronic acid.
Mol Cell Endocrinol 1982 Sep
PMID:Glycosaminoglycans in bovine cumulus-oocyte complexes: morphology and chemistry. 629 Feb 89

beta-N-Acetylglucosaminidase secreted by Entamoeba histolytica was extracted from the growth medium by affinity chromatography on CH-Sepharose 4 B coupled to p-aminophenyl-1-thio-beta-2-acetamido-2-deoxyglucopyranoside. The enzyme was further purified by isoelectric focusing, by sequential chromatography on DEAE-cellulose and Sephadex G-150, and by preparative disc gel electrophoresis. Chitobiose (betaGlcNAc1-4GlcNAc) derived from chitin as well as the oligosaccharides betaGlcNAc1-4 betaGlcUA1-3GlcNAc, betaGlcNAc1-4 betaGlcUA1-3 betaGlcAc1-4GlcUA, and betaGlcNAc1-4 betaGlc-UA1-3 betaGlcNAc1-4 betaGlcUA1-3 betaGlcNAc1-4GlcUA derived from hyaluronic acid were tested as potential physiological substrates. All these oligosaccharides are susceptible to action of beta-N-acetylglucosaminidase from E. histolytica. Under identical conditions chitobiose is cleaved 38-48 times faster than hyhyauronate oligosaccharides. No release of N-acetylglucosamine was observed when glycopeptides from ovalbumin were used as substrate. The pH optimum of hydrolase activity was 4.5 when chitobiose was used as substrate. Optimal hydrolysis of aluronate oligosaccharides was observed at pH 3.0 for trisaccharide and pH 2.0 for tetra- and hexasaccharide, respectively. Estimation of molecular weight by means of gel filtration gave values of 75 000. The isoelectric point was 5.02 beta-N-Acetylglucosaminidase from E. histolytica does not act on macromolecular chitin and hyaluronic acid.
Mol Biochem Parasitol 1983 Feb
PMID:Degradation of biogenic oligosaccharides by beta-N-acetyl-glucosaminidase secreted by Entamoeba histolytica. 630 11

The teguments of 6 and 10 day-old Hymenolepis diminuta were removed with Triton X-100 and separated into brush border and vesicular fractions by differential centrifugation. Glycosaminoglycans (GAG) isolated from these tissues and from the denuded carcass were treated with specific GAG-degrading enzymes and other chemical agents and analyzed by sodium dodecyl sulfate-polyacrylamide, agarose gel and cellulose acetate electrophoresis. Both 6 and 10 day-old worm carcasses contained chondroitin sulfate, heparin/heparan sulfate and hyaluronic acid. The 10 day-old worm brush border and vesicle fractions contained chondroitin sulfate but no heparin-like material. Colorimetric analysis showed that the carcasses of both 6 and 10 day-old worms contained uronic acid. About 98% of the detectable uronic acid of 10 day-old worms was found in the carcass, and only 2% in the brush border fraction. No uronic acid was detected in the other tegumental fractions.
Mol Biochem Parasitol 1984 Jun
PMID:Glycosaminoglycans of tegumental fractions of Hymenolepis diminuta. 643 45

Polysaccharides and other complex carbohydrates were released by proteolysis of the chloroform-methanol insoluble residue of 10 day-old worms and eggs of Hymenolepis diminuta. Gas-liquid chromatographic analysis of alditol acetate derivatives of monosaccharides released from the polysaccharides by hydrolysis revealed that in the 10 day-old worm, glucose was the most abundant sugar, followed by galactose, glucosamine, galactosamine, fucose and possibly rhamnose. Mannose was least abundant and xylose was absent. In the egg, glucose and galactose were equally abundant, followed by the same sugars found in 10 day-old worms, and xylose was present. Uronic acid was detected in both fractions by specific chemical tests. None of the saccharide material from eggs and worms was susceptible to degradation by Streptomyces hyaluronidase, chondroitinase AC, and slightly susceptible to chondroitinase ABC, as shown by electrophoretic analysis on composite 2.2% acrylamide-agarose slab gels and 4.5/12.5% polyacrylamide gels before and after enzymatic treatment. One of the gel-separable bands, however, was degradable by both nitrous acid and Flavobacterium heparinase. Both bands from eggs were degradable by nitrous acid. These results suggest that eggs contain heparin and/or heparan sulfate and perhaps dermatan sulfate and that 10 day-old worms also have these polyglycans but possibly not chondroitin sulfate or hyaluronic acid.
Mol Biochem Parasitol 1984 Jan
PMID:Characterization of polysaccharides of the eggs and adults of Hymenolepis diminuta. 653 86

The hyaluronic acid double helix, contrary to an earlier visualization, probably incorporates extensively hydrogen-bonded chains and is pinned together by carboxyl-carboxylate hydrogen bonds and water bridges. Transient interactions between stiffened chain segments provided by the formation of double-helical loops could give rise to the characteristic viscoelastic properties of hyaluronic acid solutions.
J Mol Biol 1983 Oct 05
PMID:Hyaluronic acid double helix. 663 55


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