Showing papers on "Chondroitin sulfate published in 1977"
TL;DR: After chondroitinase digestion of bovine nasal and tracheal cartilage proteoglycans, subsequent treatment withtrypsin or trypsin followed by chymotrypsin yielded two major types of polypeptide-glycosaminoglycan fragments which could be separated by Sepharose 6B chromatography.
225 citations
TL;DR: Activity levels of the chain-initiating glycosyltransferases as well as the extent of xylosylation of core protein were found to be similar in cell extracts from both culture types, and beta-xylosides did not inhibit the xylOSyltransferase reaction in cell-free studies.
183 citations
TL;DR: The synthesis of glycosaminoglycans by human skin fibroblasts derived from normal subjects, Hurler and Marfan patients before and after transformation by SV40 virus has been studied.
168 citations
TL;DR: Proteoglycan monomers were subfractionated according to buoyant density by dissociative CsCl density gradient centrifugation and it was concluded that proteoglycans vary in size mainly because of variations in the size of the chondroitin sulfate-enriched region.
167 citations
TL;DR: It is shown that increased hyaluronic acid synthesis in the embryo is correlated with the formation of the primary mesenchyme and its associated cell free spaces and the neural folds.
98 citations
TL;DR: Sedimentation equilibrium studies on the protein core of this fraction, isolated by column chromatography from chondroitinase ABC digests, suggest that its molecular weight is 2.0 x lo’ to 2.2 x 1W.
84 citations
TL;DR: Glycosaminoglycans are isolated from approximately 250 mg of aortic tissue by enzymatic degradation of tissue, followed by potassium acetate precipitation of GAGs, which are quantitated by densitometric scanning of cellulose acetate electrophoretograms.
48 citations
Journal Article•
TL;DR: C activation by polyanion-polycation interactions in the presence of CRP may be important to certain reactions of host defense and inflammation.
Abstract: Interactions between heparin and protamine previously were found to result in activation of the complement (C) system. In the present investigation, this interaction was shown to result in the binding of purified C1, and this was markedly enhanced in the presence of C-reactive protein (CRP). CRP also enhanced C consumption during heparin-protamine interactions in whole serum, and in the presence of CRP depletion of C components C1-3 was observed. Similar C1 binding and C consumption in the presence of CRP were seen upon the interaction of multiple additional polyanions including DNA, ENA, hyaluronic acid, chondroitin sulfate, and dextran sulfate with the polycations protamine sulfate and poly-L-lysine. These effects were observed with CRP concentrations well within the range found in normal human sera and considerably less than those found in most acute phase sera. We suggest, therefore, C activation by polyanion-polycation interactions in the presence of CRP may be important to certain reactions of host defense and inflammation.
45 citations
TL;DR: It is concluded that incorporation of exogenous precursors into glycosaminoglycans in the chick retina decreases relative to cell number as differentiation progresses from a period of high mitotic activity to one of tissue specialization, and that it is accompanied by a net accumulation of gly cosaminoglycan and a change in the pattern of its synthesis.
42 citations
TL;DR: Acidic glycosaminoglycans in human plasma were studied by enzymatic and electrophoretic procedures after fractionation by gel filtration on Sephadex G-100 and found to be susceptible to specific chondroitinases and streptomyces hyaluronidase.
36 citations
TL;DR: Incorporation of sulfate into sulfated proteoglycans by isolated chicken chondrocytes was inhibited up to 74% by transformation with the Rous sarcoma virus, and a similar inhibitory effect was observed on acetate incorporation into chondroitin sulfate.
Abstract: Incorporation of sulfate into sulfated proteoglycans by isolated chicken chondrocytes was inhibited up to 74% by transformation with the Rous sarcoma virus, and a similar inhibitory effect was observed on acetate incorporation into chondroitin sulfate. Slower sedimenting sulfated proteoglycans appear after the viral transformation. The ratio of chondroitin 4-sulfate to chondroitin 6-sulfate in these slower sedimenting sulfated proteoglycans was different from that of normal chondrocytes, but the chain lengths of sulfated glycosaminoglycans produced by normal chondrocytes and transformed chondrocytes were not significantly different. Chondrocytes were also infected with a temperature-sensitive mutant of RSV, ts LA24, which has a temperature-sensitive lesion in the transforming gene. Hyaluronic acid production by these cells was increased, and the slower sedimenting sulfated proteoglycan was produced only at the permissive temperature.
TL;DR: Although mass cultures contain some non-muscle cell types, susceptibility of the autoradiographic grains associated with cells in muscle clones to testicular hyaluronidase digestion suggests that at least some glycosaminoglycans (chondroitin sulfate, in particular) were synthesized by muscle cells.
Abstract: Day 3 mass muscle cultures incorporated 35SO4= and 3H· glucosamine into specific glycosaminoglycans that were identified by a variety of methods (enzyme susceptibility, nitrous acid degradation, electrophoresis, thin layer chromatography). More than 75% of the newly synthesized glycosaminoglycans appeared in the medium. In the medium, the glycosaminoglycans consisted of primarily hyaluronic acid (∼35%) and chondroitin sulfate (∼40%) with smaller amounts of chondroitin (∼15%) and heparan sulfate (∼10%). Among the glycosaminoglycans that remained in the cell layer, chondroitin sulfate (∼45%) and heparan sulfate (20-40%) predominated and only a small amount of chondroitin (∼15%) and hyaluronic acid (∼5%) were present. In both the medium and cell layer, greater than 80% of the chondroitin sulfate was chondroitin-6-sulfate. Although mass cultures contain some non-muscle cell types which undoubtedly contributed to total glycosaminoglycan synthesis, susceptibility of the autoradiographic grains associated with cells in muscle clones to testicular hyaluronidase digestion suggests that at least some glycosaminoglycans (chondroitin sulfate, in particular) were synthesized by muscle cells.
Transmission electron microscopy revealed the presence of ruthenium red staining globular units along the surfaces of myotubes. Destruction of the integrity of surface associated ruthenium red stained material by both Streptomyces hyaluronidase and testicular hyaluronidase suggests that both hyaluronic acid and chondroitin sulfate are present on muscle cell surfaces.
TL;DR: 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 hyaluonic acid is not a structural backbone for glycosamine complexes of the external cell surface.
Abstract: 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.
TL;DR: It is suggested that independent sorting of these relatively few glycopeptides can result in qualitative surface differences and Morphological state (epithelial or mesenchymal) and differentiated state may be related to these differences as well as to quantitative differences in surface glycosaminoglycans.
Abstract: Anionic glycopeptides and glycosaminoglycans synthesized by embryonic neural tube (epithelium) and neural crest (mesenchymal outgrowth) developing in vitro were examined. The profile of surface glycopeptides is relatively simple (two major ones for neural crest; four major ones for neural tube). There is one major glycopeptide found in the medium which is present only in trace amounts on the cells. Both hyaluronate and chondroitin sulfate are synthesized. Hyaluronate is predominantly cell associated; chondroitin sulfate is found predominantly in the medium. It is suggested that independent sorting of these relatively few glycopeptides can result in qualitative surface differences. Morphological state (epithelial or mesenchymal) and differentiated state may be related to these differences as well as to quantitative differences in surface glycosaminoglycans.
TL;DR: Treatment with (+)-catechin produced an improvement, in both patients, of the biochemical pattern of collagen and GAGs and a shift of the cellular activity and of the matrix morphology towards normality was observed in the investigated cartilage of the less severely affected patient.
Abstract: Two patients affected with two different forms of Osteogenesis Imperfecta were examined in order to study collagen and glycosaminoglycans (GAGs) in skin and iliac crest cartilage. A sharp decrease of the galactosamine to glucosa-mine ratio due to a reduced content of chondroitin sulfate was evidenced in both patients. Moreover the structure of proteoglycans appeared altered, this being more evident in the severe form of the disease. Morphological examination in light and electron microscopy of cartilage of the less severely diseased patient showed that GAGs in the extracellular matrix did not present regular connection with collagen fibers. Chondrocytes, elongated and disorderly scattered, showed large lipidic inclusions and, on histochemical basis, were devoid of UDPG dehydrogenase activity. Treatment with (+)-catechin produced an improvement, in both patients, of the biochemical pattern of collagen and GAGs. Similarly a shift of the cellular activity and of the matrix morphology towards normality was ob...
TL;DR: It was found that the estrogen treatment specifically increases the fast-migrating sialoglycoprotein level in the luminal epithelia and results in the appearance of it in the uterine fluid.
TL;DR: Data suggest that UDP-xylose concentration in chondrocytes is regulated by substrate activation and product inhibition of UDP-glucuronate carboxy-lyase, and a common mechanism governs induction of glycosyltransferases and of UDP's allosterically activated and inhibited status.
Journal Article•
TL;DR: Human and rat decalcified bone matrix preparations were shown to be active in inducing cartilage formation by subcutaneous implantation in the rat and it is suggested that this approach to the culture of connective tissue cells will allow a more precise definition of the early steps of connectives tissue differentiation.
TL;DR: It is postulated that induction of platelet-cartilage interaction may eventuate in cartilage repair and some degree of cartilage healing may take place after initiation of an inflammatory response.
Abstract: During studies concerned with the platelet-collagen interaction, it was observed that platelets did not adhere to bovine or human articular cartilage and that cartilage did not induce platelet aggregation in vivo or in vitro. To study the mechanism responsible for this observation, the role of proteoglycans was examined. Purified cartilage collagen proved to be fully active as a platelet aggregant. Addition of small amounts of proteoglycan subunit (PGS) blocked platelet aggregation, whereas chondroitin sulfate, a major glycosaminoglycan component of cartilage matrix, impaired platelet aggregation only at concentrations which resulted in a marked increase in viscosity. Moreover, PGS abolished aggregation of platelets by polylysine but did not prevent aggregation by ADP, suggesting that PGS may block strategically placed lysine sites on the collagen molecule. Treatment of fresh articular cartilage with proteolytic enzymes rendered the tissue active as a platelet aggregant. In vivo experiments demonstrated that surgical scarification of rabbit articular cartilage does not result in adhesion of autologous platelets. Treatment of rabbit knee joints with intraarticular trypsin 1 wk before the injection of blood resulted in adhesion and aggregation of platelets on the surface of the lesions. Since there is evidence from other studies that some degree of cartilage healing may take place after initiation of an inflammatory response, it is postulated that induction of platelet-cartilage interaction may eventuate in cartilage repair.
TL;DR: An hypothesis is proposed to account for the interaction between the cell and its extracellular matrix macromolecules and it is proposed that external hyaluronic acid inhibits chondroitin sulfate production.
Abstract: Extracellular matrix macromolecules are capable of complex interactions. Recent studies demonstrate that extracellular matrix macromolecules can regulate the synthesis of chrondroitin sulfate by chondrocytes. Extracellular chondroitin sulfate stimulates chondroitin sulfate synthesis, whereas external hyaluronic acid inhibits chondroitin sulfate production. Other matrix macromolecule-cell interactions are reviewed. An hypothesis is proposed to account for the interaction between the cell and its extracellular matrix macromolecules.
TL;DR: Until proteases can be rigorously excluded from enzyme preparations used to degrade the proteoglycan subunit, it will not be possible to determine whether it consists of a single or several different core proteins.
Abstract: The basic subunit of cartilage proteoglycan consists of multiple glycosaminoglycan chains covalently attached to a core protein. It is unclear as to whether there is a single core protein or multiple different core proteins, since previous studies using either chondroitinase or testicular hyaluronidase to enzymatically remove chondroitin sulfate side chains from the proteoglycan subunit have yielded conflicting results. In the present study, a chondroitinase-produced core protein preparation isolated as a single peak on Sepharose gel chromatography was found to contain at least two immunologically distinct components. Hyaluronidase-produced core protein from the same proteoglycan subunit fraction was found to contain multiple components nearly all of which were smaller than the components in the chondroitinase digest. A possible explanation of these findings is that they resulted from proteolytic degradation of the core protein in the course of the enzymatic removal of its chondroitin sulfate. The presence of small amounts of protease contaminants in several commercial chondroitinase and hyaluronidase preparations was detected by an extremely sensitive radioassay. Until proteases can be rigorously excluded from enzyme preparations used to degrade the proteoglycan subunit, it will not be possible to determine whether it consists of a single or several different core proteins.
TL;DR: The results indicated that human gastric AGAG consisted of, in the order of amount, heparan sulfates, dermatan sulfate, hyaluronic acid, chondroitin-4-sulfate, ChondroitIn-6-Sulfate and presumably oversulfated chondDetroitin sulfate.
Abstract: The acidic glycosaminoglycans (AGAG) in normal portions of human gastric tissue were separated by electrophoresis in 3 buffer systems. Paper chromatographic separation of the constitutional disaccharide units by digestion of chondroitin sulfates (CS) with chondroitinase-ABC and chondroitinase-AC was carried out after fractionation of CS by ion-exchange resin column chromatography. Thin-layer chromatography of hexosamines and other biochemical analysis were also performed. The presence of hyaluronic acid in the gastric tissue was substantiated by the enzymatic susceptibility to streptomyces hyaluronidase. The results indicated that human gastric AGAG consisted of, in the order of amount, heparan sulfates, dermatan sulfate, hyaluronic acid, chondroitin-4-sulfate, chondroitin-6-sulfate and presumably oversulfated chondroitin sulfate.
TL;DR: It is shown by [3H]NaBH4 reduction and subsequent identification of the 3H-labelled sugar alcohols that heparan sulfate is degraded in all organs by at least two endoglycosidases, an endoglucuronidase and an endglucosaminidase, to fragments of low molecular weight.
Abstract: The characterization of intracellularly stored glycosaminoglycans from organs of a patient suffering from mucopolysaccharidosis III A (Sanfilippo A disease) is described. Both heparan sulfate and galactosamine-containing glycosaminoglycans (chondroitin sulfate, dermatan sulfate) are accumulated in the liver, whereas in the other organs (spleen, kidney, heart, cerebrum, cerebellum) heparan sulfate is almost the only glycosaminoglycan stored. It is shown by [3H]NaBH4 reduction and subsequent identification of the 3H-labelled sugar alcohols that heparan sulfate is degraded in all organs by at least two endoglycosidases, an endoglucuronidase and an endoglucosaminidase, to fragments of low molecular weight (Mr approximately 2 000-6 600).
TL;DR: In this paper, two-dimensional electrophoresis on cellulose acetate membranes was used to study the mechanism of repair of myocardium after infarction, showing that the repair process of infarcted myocardia is very similar to that occurring during healing of hepatic fibrosis and damaged arterial walls.
TL;DR: Although the cartilage proteoglycan macromolecules were heterogeneous, a certain regularity was observed with respect to the distribution of sulfate and the degree of sulfation in the chondroitin sulfate chains of the proteoglycans.
Abstract: Proteoglycans were extracted from bovine articular cartilage with guanidine-HCl and fractionated in cesium chloride density gradients by equilibrium ultracentrifugation. The acidic glycosaminoglycan (AGAG) components were then determined enzymatically with chondroitinase-ABC and streptomyces hyaluronidase. Under associative and dissociative conditions, the distribution of the AGAG components was as follows: the ratio of 4-sulfated disaccharide units to total AGAG increased with decreasing density gradients whereas that of 6-sulfated disaccharide units to total AGAG increased with increasing density gradients. The ratio of disulfated disaccharide units to total AGAG increased somewhat with decreasing density gradients whereas that of non-sulfated disaccharide units tended to decrease. Although the cartilage proteoglycan macromolecules were heterogeneous, a certain regularity was observed with respect to the distribution of sulfate and the degree of sulfation in the chondroitin sulfate chains of the proteog...
Journal Article•
TL;DR: Glycosaminoglycans were determined in the blood plasma of rabbits as a function of time after intravenous, intramuscular and intragastric administration of chondroitin sulfate preparation.
Abstract: Glycosaminoglycans were determined in the blood plasma of rabbits as a function of time after intravenous, intramuscular and intragastric administration of chondroitin sulfate preparation. After intravenous administration two phases of chondoitin sulfate elimination from the blood with T1/2 131-135 min and 8.5-9.5 min, respectively were observed. The level of the blood plasma glycosaminoglycans did not increase after intragastric introduction of the chondroitin sulfate preparation. Excretion of glycoaminoglycans in the urine was also determined.
TL;DR: The mucopolysaccharides produced by B16 mouse melanoma cells have been isolated from the spent media in which the cells were grown in the presence of 2-amino-2-deoxy-D-glucose-t and [35S]-sulfate and confirmed their similarity to chondroitin-4-sulfates from cartilage.
TL;DR: Of the natural glycosaminoglycans tested, only heparin was a potent inhibitor of poly (U)-directed polyphenylalanine synthesis by rat liver ribosomes (50% inhibition at 10 mug/ml); a chemically oversulfated chondroitin sulfate was twice as effective, and another synthetic polyanion, sodium pentosan polysulfate was ten times more effective than heparins.
Abstract: Of the natural glycosaminoglycans tested, only heparin was a potent inhibitor of poly (U)-directed polyphenylalanine synthesis by rat liver ribosomes (50% inhibition at 10 mug/ml). A chemically oversulfated chondroitin sulfate was twice as effective, and another synthetic polyanion, sodium pentosan polysulfate was ten times more effective than heparin. Chondroitin-4,6-sulfate was inhibitory at very high concentrations (15 mg/ml) and heparan sulfate at concentrations above 100 mug/ml. The compounds interfere with the formation of ternary complex consisting of the ribosome, poly(U) and phenylalanyl-tRNA. The inhibitors prevented the attachment of the mRNA to the ribosome, probably by competition with poly(U) for the ribosomal binding site of mRNA. However, they were ineffective in doing so once phenylalanyl-tRNA has bound to the ribosomepoly(U) complex. Sucrose gradient analysis in presence of the inhibitors revealed a selective effect on the sedimentation of the small ribosomal subunit; the large subunit was unaltered. This effect, however, was dependent on the concentration of magnesium. In contrast to Escherichia coli ribosomes, no binding of the inhibitors to the particles could be demonstrated.
TL;DR: In an attempt to understand the nature of the nonextractable GAG and their relation to fibrous proteins, bovine aorta was selectively hydrolyzed with elastase and collagenase and GAG were isolated from the hydrolyzates, characterized and quantitated.
Abstract: At the present time little is known about the biologic role of glycosaminoglycans (GAG) in cardiovascular tissues or the role they play with other connective tissue components in cardiovascular tissues Elucidation of the interactions between GAG and the fibrous proteins would aid in the understanding of the molecular organization within cardiovascular structure In earlier studies of the aorta (1) proteoglycans (GAG with covalently bound proteins) were extracted by several dissociative solvents These studies indicate that about half of the total uronic acid contained in the tissue was isolated The uronic acid material extracted by dissociative methods is part of a proteoglycan containing both chondroitin sulfates (CS) and dermatan sulfate (DS) No heparan sulfate (HS) was obtained although aorta contains considerable amount of this GAG In an attempt to understand the nature of the nonextractable GAG and their relation to fibrous proteins, bovine aorta was selectively hydrolyzed with elastase and collagenase and GAG were isolated from the hydrolyzates, characterized and quantitated
TL;DR: The chromatographic pattern of the urinary chondroitin sulfate fraction in patients with angiosarcoma of the liver differed from those of controls in that there was a relative increase in the total amount of uronic acid in a hyaluronidase‐resistant fraction and a decrease in a fraction susceptible to hyaluronic acid digestion.
Abstract: Glycosaminoglycans extracted from 24-hour urine specimens from patients with hepatic angiosarcoma and from normal/controls were separated as cetylpyridinium complexes into “hyaluronic acid,” “chondroitin sulfate,” and “heparin” fractions, then further separated and characterized by anion-exchange chromatography and hyaluronidase susceptibility. The chromatographic pattern of the urinary chondroitin sulfate fraction in patients with angiosarcoma of the liver differed from those of controls in that there was a relative increase in the total amount of uronic acid in a hyaluronidase-resistant fraction and a decrease in a fraction susceptible to hyaluronidase digestion. These changes appeared to become more pronounced with advancing disease. Chromatographic patterns and determinations of hyaluronidase susceptibility indicated that the resistant fraction was heparan sulfate and that the susceptible fraction was chondroitin-4-sulfate and/or chondroitin-6-sulfate. Cancer 40:3050-3053, 1977.