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Showing papers on "Keratan sulfate published in 1986"


Journal ArticleDOI
TL;DR: The chondroitin sulfate proteoglycan of rat brain was digested with Pronase, and after removal of glycosaminoglycans, the resulting glycopeptides were treated with alkaline borohydride to release O-glycosidically linked oligosaccharides and the combined decrease in mannose and N-acetylgalactosamine was very close to the observed destruction of serine + threonine.

154 citations


Journal ArticleDOI
TL;DR: Antibodies to corneal keratan sulfate proteoglycan (KSPG) were used to characterize the pattern of KSPG accumulation during differentiation of neural crest cells in the stroma of embryonic chick cornea, suggesting a modulation of K SPG primary structure late in development and after hatching.

115 citations


Journal ArticleDOI
TL;DR: The results of these studies suggest that articular chondrocytes have an inherent program that determines the quality of proteoglycans synthesized at different ages.

71 citations


Journal ArticleDOI
TL;DR: It is apparent from this study that certain GAG species are incorporated into the structure of the stone and they may be intimately related to stone development and growth.

69 citations


Journal ArticleDOI
TL;DR: Sulfated keratan sulfate was not detected in the serum of 16 patients with macular corneal dystrophy, but was present at normal levels in 66 patients with other cornea diseases, and this assay should prove useful in the diagnosis of macular Corneal Dystrophy.

61 citations


Journal ArticleDOI
01 Dec 1986
TL;DR: Keratan sulfate in the serum appears to be derived predominantly from the normal turnover of cartilage, and these studies strongly suggest that the defect in keratan sulfates synthesis in MCD is not restricted to corneal cells and that M CD is one manifestation of a systemic disorder of keratan sulfur.
Abstract: An ELISA assay using a monoclonal antibody (ET-4-A-4) that recognizes a sulfated carbohydrate epitope in both keratan sulfate type I (corneal) and type II (skeletal) was employed to quantify keratan sulfate in serum and corneal tissue from patients with macular corneal dystrophy (MCD). This assay disclosed significant quantities of keratan sulfate in the serum in 45 healthy individuals (251 +/- 78 ng/ml), and in 66 patients with various corneal diseases (273 +/- 101 ng/ml). In contrast keratan sulfate was not detected (less than 2 ng/ml) in the serum of 16 patients with histopathologically confirmed MCD. Keratan sulfate was also detected in extracts of normal corneas and corneal tissue with a variety of pathologic conditions, but was virtually absent in corneal tissue from five patients with MCD. In corneas with MCD the chondroitin sulfate/keratan sulfate ratio was considerably higher than that of all normal and pathologic corneas studied. Since keratan sulfate in the serum appears to be derived predominantly from the normal turnover of cartilage these studies strongly suggest that the defect in keratan sulfate synthesis in MCD is not restricted to corneal cells and that MCD is one manifestation of a systemic disorder of keratan sulfate. The cartilage changes, however, do not have clinical significance. Moreover, since keratan sulfate can be detected in the blood of newborns it should be possible to diagnose MCD prior to corneal opacification.

46 citations


Journal ArticleDOI
TL;DR: The results clearly show that sulphate residues are essential components of the antigenic determinant(s) recognised by three monoclonal antibodies to keratan sulphate, but they mask the i antigen activity of the linear poly-(N-acetyllactosamine) backbones of this glycosaminoglycan.
Abstract: Conditions were established for desulphation of hexa-, octa-, deca- and larger oligosaccharides derived from corneal keratan sulphate after treatment with endo-beta-galactosidase. The antigenicities of the desulphated oligosaccharides were compared with those of the native oligosaccharides in chromatogram binding, plastic-plate binding or inhibition of binding assays using a novel microimmunochemical approach with oligosaccharide-lipid conjugates (neoglycolipids). The results clearly show that sulphate residues are essential components of the antigenic determinant(s) recognised by three monoclonal antibodies to keratan sulphate, 5-D-4, 1-B-4 and MZ15, but they mask the i antigen activity of the linear poly-(N-acetyllactosamine) backbones of this glycosaminoglycan. Immunochemical assays, before and after beta-N-acetylglucosaminidase treatment of desulphated linear hexa-, octa- and decasaccharides derived from keratan sulphate, indicate that for reaction with one anti-i antibody, Den, there is an absolute requirement for the non-reducing beta-galactosyl residue of the i antigen structure to be in the terminal position, but with a second anti-i antibody, Tho, there is in addition some reactivity with the i antigen structure having an N-acetylglucosamine residue at the non-reducing end. The chromatographic properties after desulphation or nitrosation of a minor keratan sulphate oligosaccharide (a dodecasaccharide), which reacts especially well with antibody 5-D-4, have provided the first evidence for the presence of glucosamine residues that may be N-sulphated in corneal keratan sulphate.

40 citations


Journal ArticleDOI
H Inoue, K Otsu, M Yoneda, K Kimata, S Suzuki, Y Nakanishi 
TL;DR: The results suggest that the appearance of the sulfotransferases in serum is not a fortuitous event due to nonspecific cell death, but the result of an elaborate mechanism for enzyme secretion by a cell or tissue system.

34 citations


Journal ArticleDOI
TL;DR: It is suggested that the differences found between first and second extractions of cartilage, between upper and lower layers of Cartilage, and between knee and hip cartilage are caused by variations in the relative amount of nonaggregating proteoglycans and/or variations in proteoglycan size.
Abstract: Proteoglycans (A1 fractions) were extracted with 4M guanidine hydrochloride (GuHCl) from human articular cartilage samples of a wide age range Distinctions were made between hip and knee, and upper and lower layers The residues of these extractions were digested with purified collagenase, and a second extraction with 4M GuHCl was performed, which yielded appreciable amounts of proteoglycans When pro-teoglycans from second extractions were compared with those from first extractions, the following changes were observed: an increase in chondroitin sulfate; a relative decrease in keratan sulfate; a decrease in protein content; and a decrease in the ratio of chondroitin 6-sulfate to chondroitin 4-sulfate The same changes were found when nonaggregating proteoglycans were compared with proteoglycan aggregates, when proteoglycans from young cartilage were compared with those from mature cartilage, when proteoglycans from knee cartilage were compared with those from hip cartilage, and when proteoglycans from upper layers of cartilage were compared with those from deeper layers It is suggested that the differences found between first and second extractions of cartilage, between upper and lower layers of cartilage, and between knee and hip cartilage are caused by variations in the relative amount of nonaggregating proteoglycans and/or variations in proteoglycan size

26 citations


Journal ArticleDOI
TL;DR: The results suggest that although KS synthesis starts at very early stages of fetal development, there are progressive changes in its antigenic structure in specific regions of the cornea and conjunctiva during corneal development.
Abstract: Twenty-six monoclonal antibodies (MAbs) developed against rabbit corneal proteokeratan sulfate (PKS), were used to evaluate immunohistochemically the ocular distribution of PKS during prenatal and early postnatal development in rabbits These MAbs were directed against epitopes located in the keratan sulfate (KS) chains of the proteoglycan (SundarRaj et al, 1985) Staining of cryostat sections of the eyes was carried out using an indirect peroxidase-conjugated technique Only one of the MAbs reacted with the presumptive corneal region at day 13 or 16 of fetal development By day 20, more MAbs reacted with the corneal stroma There were distinct differences, however, in the distribution of the epitopes recognized by the various MAbs A few of them stained only the posterior region of the cornea, whereas others showed a decreasing staining gradient from the posterior to the anterior region By day 24, all of the MAbs reacted with the corneal stroma, but some reacted also with the limbal region and with the conjunctival stromal matrix One MAb also reacted with the conjunctival epithelial layer, but only at this stage of development Conjunctival staining was more intense at day 28 of fetal development and at day 2 postnatally KS was not detectable in the conjunctiva of adult rabbits with any of the MABs These results suggest that although KS synthesis starts at very early stages of fetal development, there are progressive changes in its antigenic structure in specific regions of the cornea and conjunctiva during corneal development

25 citations


Journal ArticleDOI
TL;DR: Keratan sulphate was extracted from a shark/whale cartilage preparation and examined by 400 MHz 1H- and 100 MHz 13C-n.r.m. spectroscopy, showing consistent with an N-acetyl-lactosamine repeating unit that is predominantly sulphated at C-6 of both galactose and N- acetylglucosamine.
Abstract: Keratan sulphate was extracted from a shark/whale cartilage preparation and examined by 400 MHz 1H- and 100 MHz 13C-n.m.r. spectroscopy. Assignment of the majority of the resonances was facilitated by two-dimensional 13C-1H correlation by using a modified COLOC procedure and a COSY-45 experiment. The spectra are consistent with an N-acetyl-lactosamine repeating unit that is predominantly sulphated at C-6 of both galactose and N-acetylglucosamine. Gel chromatography of a keratanase digest of the shark keratan sulphate confirmed the high degree of galactose sulphation.

Journal ArticleDOI
TL;DR: Findings clearly show that the sulfation on the N-acetylglucosamine adjacent to galactose in the lactosaminoglycans is essential for expression of the Pseudomonas enzyme, but not for that of the Flavobacterium or Escherichia enzyme.
Abstract: The substrate specificity of endo-beta-galactosidase of Pseudomonas sp. was found to differ from that of Flavobacterium keratolyticus or Escherichia freundii, based on the following experimental results. The endo-beta-galactosidases from these three bacteria released 6-O-sulfo-GlcNAc beta 1-3Gal as one of the major products from keratan sulfates from different sources. In addition to the sulfated disaccharide, Flavobacterium and Escherichia enzymes produced GlcNAc beta 1-3Gal, which is also an integral repeating unit of keratan sulfate, whereas the Pseudomonas enzyme did not release any non-sulfated disaccharide. Tetrasaccharides were prepared from the teleost skin keratan sulfate by digestion with Pseudomonas enzyme followed by gel filtration on Sephadex G-50 chromatography. A part of the tetrasaccharide fraction was hydrolyzed by Flavobacterium enzyme to produce 6-O-sulfo-GlcNAc beta 1-3Gal and GlcNAc beta 1-3Gal, whereas the fraction was completely resistant to retreatment with the Pseudomonas enzyme. Endo-beta-galactosidases from F. keratolyticus and E. freundii hydrolyzed the internal beta-1,4-galactosyl linkage of various neolacto-type glycosphingolipids to produce glucosylceramides. However, these glycosphingolipids were completely resistant to the Pseudomonas enzyme. These findings clearly show that the sulfation on the N-acetylglucosamine adjacent to galactose in the lactosaminoglycans is essential for expression of the Pseudomonas enzyme, but not for that of the Flavobacterium or Escherichia enzyme.

Journal ArticleDOI
TL;DR: It appears that the role of the squid sulfotransferase is to synthesize so-called chondroitin sulfate E where over 50% of the interior hexosamine units are 4,6-bis-sulfated.

Journal ArticleDOI
TL;DR: Results suggest that in cultured chondroblasts KS:CS-PG and Type II procollagen are differentially distributed both in organelles and in the extracellular matrix, and that different proteoglycan types may occupy distinct subcompartments in trans Golgi.
Abstract: The mechanisms of synthesis and intracellular routing of the various cartilage matrix macromolecules are still unclear. We have studied this problem in cultured chondroblasts at the ultrastructural level using (i) monospecific antibodies against the core protein of the keratan sulfate/chondroitin sulfate-rich cartilage proteoglycan (KS:CS-PG) or Type II procollagen, and (ii) cuprolinic blue, a cationic dye that binds to the glycosaminoglycan chains of proteoglycans. Intracellularly, the proteoglycan antibodies localized KS:CS-PG and its precursors primarily in the Golgi complex and secretory vesicles. In contrast, the bulk of Type II procollagen was found within the rough endoplasmic reticulum (ER). While devoid of collagen, the extracellular matrix was rich in KS:CS-PG molecules some of which studded the chondroblast plasmalemma. Cuprolinic blue staining indicated that the proteoglycans present in the Golgi complex fell into a predominant class of large proteoglycans, probably representing KS:CS-PG, and a minor class of smaller proteoglycans. Groups of these divergent proteoglycans often occupied distinct Golgi subcompartments; moreover, single large proteoglycans appeared to align along the luminal surface of Golgi cisternae and secretory vesicles. These results suggest that in cultured chondroblasts KS:CS-PG and Type II procollagen are differentially distributed both in organelles and in the extracellular matrix, and that different proteoglycan types may occupy distinct subcompartments in trans Golgi.

Journal ArticleDOI
TL;DR: It is postulated that most species of glycosaminoglycans restricted non-specific fluid phase complement consumption induced by LIS, an effect which conserved complement and thereby enhanced the subsequent residual serum C mediated hemolytic activity.

Journal ArticleDOI
TL;DR: Results indicates that the enzyme catalyses the hydrolysis of beta-D-galactose-(1----4)-N-acetylglucosamine linkages, and it was shown that this monosulfated disaccharide inhibits the particulate keratan sulfate endoglycosidase.
Abstract: Four constitutive enzymes, capable of degrading keratan sulfate, were isolated from Pseudomonas sp.: a particulate endoglycosidase, a soluble endoglycosidase, a soluble exo-beta-D-galactosidase and a soluble exo-beta-D-N-acetylglucosaminidase. The endoglycosidases were shown to act only upon keratan sulfate forming beta-D-2-acetamido-2-deoxy-6-O-sulfoglucosyl-(1----3)-D-galactose, as the main product. This results indicates that the enzyme catalyses the hydrolysis of beta-D-galactose-(1----4)-N-acetylglucosamine linkages. It was also shown that this monosulfated disaccharide inhibits the particulate keratan sulfate endoglycosidase. The bovine nucleus pulposus keratan sulfate is depolymerized at a lower rate and extent when compared to the corneal keratan sulfate. The soluble endoglycosidase is very labile, in contrast to the particulate enzyme, which has been stored at -20 degrees C or at 4 degrees C for at least 12 months with no loss in activity. The particulate endoglycosidase and the soluble exo-beta-D-galactosidase and exo-beta-D-N-acetylglucosaminidase are induced when the bacteria is grown in adaptative media containing either 0.1% keratan sulfate or 0.1% chondroitin sulfate. Furthermore, particulate forms of the exoenzymes were detected. The soluble endoglycosidase specific activity, in contrast, is approximately the same in extracts of cells grown in glucose, keratan sulfate or chondroitin sulfate. A chondroitin sulfate lyase was also identified in the soluble extracts of Pseudomonas sp. cells. This enzyme depolymerizes chondroitin 4-sulfate, chondroitin 6-sulfate and hyaluronic acid forming unsaturated disaccharides as main products. It is also active upon the glucuronic-acid-containing regions of the dermatan sulfate molecules. The properties of the soluble enzymes, further purified by ion-exchange chromatography, and of the particulate keratan sulfate endoglycosidase are presented.

Journal ArticleDOI
TL;DR: The glycosaminoglycans of the myocardium of an individual who died of acute carbon monoxide poisoning were quantified by two-dimensional electrophoresis on cellulose acetate membranes and showed the presence of two unusual oversulfated chondroitin sulfates (chondroit in sulfates D and E).

Journal ArticleDOI
TL;DR: The effects of digestion with enzymes upon the histochemical reactions of the cartilage tissues indicated that hyaluronic acid exists as a glycoprotein link factor of proteoglycan subunits, forming proteogly can aggregates in tissues.

Journal ArticleDOI
TL;DR: The hydrodynamic size of proteoglycan subunits and the tissue concentration of total glycosaminoglycans decreased with increasing grade of malignancy and the molecular weight of keratan sulfate with increasing degree ofmalignancy was not observed.

Journal ArticleDOI
TL;DR: Results show predictable and physiologically regulated variation in serum keratan sulfate levels which correlate directly with specific phases of the antler regeneration cycle, and provide additional support for the assertion that measurements of keratan sulphate levels in serum can provide useful information about changes in cartilage metabolism in normal as well as diseased states.
Abstract: A sensitive and highly specific enzymelinked immunosorbent assay with an inhibition step was used to monitor the concentration of keratan sulfate, a cartilage-related glycosaminoglycan, in the serum of three adult male deer. During the course of one complete annual antler regeneration cycle, keratan sulfate levels were found to fluctuate predictably in relation to the growth and maturation of the antlers: levels are substantially elevated during the growth phase and drop precipitously when growth ceases and the antlers become fully mineralized. In addition, an unanticipated elevation of serum keratan sulfate was observed in early spring prior to casting of the preceding year's antlers and the initiation of regrowth. This suggests that changes in cartilage metabolism occur concomitantly, with this phase of the antlerogenic cycle. These results show predictable and physiologically regulated variation in serum keratan sulfate levels which correlate directly with specific phases of the antler regeneration cycle. Furthermore, the findings provide additional support for the assertion that measurements of keratan sulfate levels in serum can provide useful information about changes in cartilage metabolism in normal as well as diseased states.

Journal ArticleDOI
A. Ando1, I. Ando1
TL;DR: In this article, the structure of 67Ga-binding acid mucopolysaccharide in tumor tissues was determined from measuring neutral saccharides in the structure, and it was determined that the principal acid acid acid in the tumor was either keratan sulfate and/or keratan polysulfate.
Abstract: The present study was undertaken to determine the structure of67Ga-binding acid mucopolysaccharide in tumor tissues. It was determined from measuring neutral saccharide in the structure that the principal67Ga-binding acid mucopolysaccharide in the tumor was keratan sulfate and/or keratan polysulfate. On the other hand, it was clarified from the results of mucopolysaccharase treatment that the main67Ga-binding acid mucopolysaccharide in tumor was not keratan sulfate, heparan sulfate, heparin, nor chondroitin sulfate A, B, or C. Based on the present results, it was deduced that the main67Ga-binding acid mucopolysaccharide in tumor is keratan polysulfate and that this acid mucopolysaccharide plays the most important role in tumor accumuation of67Ga.

Journal ArticleDOI
TL;DR: The antigenic determinants of A1D1 for cellmediated immunity reside not in the glycosaminoglyc... but in the chondroitin sulfate and keratan sulfate antigens.
Abstract: The immunization of guinea pigs with 3 mg of bovine nasal cartilage proteoglycan, A1D1, in complete Freund's adjuvant (CFA) induced a delayed-type hypersensitivity (DTH) and a macrophage migration inhibition (MI) activity of peritoneal exudate cells (PEC). To assess the antigenic determinants of A1D1 in the DTH response, we examined the DTH response of guinea pigs immunized with A1D1 in CFA, using five different kinds of antigens: A1D1 digested with chondroitinase ABC, with trypsin, or with pronase, as well as authentic chondroitin sulfate and keratan sulfate. A1D1 digested with chondroitinase ABC or trypsin produced DTH responses and an MI activity on PEC. On the other hand, pronase digestion of A1D1 significantly suppressed both the DTH response and MI activity on the PEC. Authentic chondroitin sulfate and keratan sulfate, however, induced neither DTH responses nor MI activity. We have therefore concluded that the antigenic determinants of A1D1 for cellmediated immunity reside not in the glycosaminoglyc...