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The Journal of Biological Chemistry Jun 1982Previous studies on scleral proteoglycans (proteodermatan sulfate) using light scattering and ultracentrifugation techniques have shown that the molecules form...
Previous studies on scleral proteoglycans (proteodermatan sulfate) using light scattering and ultracentrifugation techniques have shown that the molecules form aggregates in 0.15 M NaCl (Cöster, L., Fransson, L.-A., Sheehan, J. K., Nieduszynski, I. A., and Phelps, C. F. (1981) Biochem. J., 197, 483-490). Aggregation was not promoted by hyaluronate but addition of free scleral dermatan sulfate chains enhanced multimerization. To investigate the possibility that scleral proteoglycans interact via their dermatan sulfate side chains, we have adopted an affinity chromatography procedure where binding of proteoglycans to various dermatan sulfate-agaroses may be studied. The evidence for an interaction between the side chains of the macromolecules and the immobilized dermatan sulfates are as follows: (a) the dermatan sulfate chains released from the proteoglycan by proteolysis display affinity for dermatan sulfate-agarose, (b) a significant proportion of the [3H]acetylated proteoglycans that were bound to the dermatan sulfate gel can be displaced by eluting with a solution of dermatan sulfate chains, (c) selective periodate oxidation of L-iduronate in the dermatan sulfate chains of the proteoglycans abolishes the affinity, (d) the core protein prepared by chondroitinase ABC digestion of the proteoglycan does not bind to dermatan sulfate-agarose, and (e) binding is retained after reduction-alkylation of the protein core. Furthermore, free [3H]dermatan sulfate chains co-elute with the proteoglycan upon gel filtration in 0.2 M NaCl.
Topics: Animals; Cattle; Chondroitin; Chromatography, Affinity; Dermatan Sulfate; Macromolecular Substances; Molecular Weight; Proteoglycans; Sclera; Uronic Acids
PubMed: 7076674
DOI: No ID Found -
Analytical Biochemistry Sep 1996A chemical method for the determination of dermatan sulfate (DS) and oversulfated dermatan sulfate has been developed and applied to the pharmacokinetic study of these...
Simultaneous determination of dermatan sulfate and oversulfated dermatan sulfate in plasma by high-performance liquid chromatography with postcolumn fluorescence derivatization.
A chemical method for the determination of dermatan sulfate (DS) and oversulfated dermatan sulfate has been developed and applied to the pharmacokinetic study of these polysaccharides in experimental animals. The analytical procedure includes a simple preparation step of administered DS and oversulfated DS from blood plasma, HPLC for the separation and detection of DS and oversulfated DS using an Asahipak NH2P-50 column, fluorometric reaction of the polysaccharides with guanidine in a strong alkaline medium. DS and oversulfated DS were extracted from plasma by treating it with proteinase to remove plasma proteins and recovered with endogenous plasma glycosaminoglycans by ethanol precipitation. Finally, DS and oversulfated DS were analyzed by fluorometric HPLC. The detection limits of DS and oversulfated DS were 10 and 20 ng, respectively. Furthermore, we demonstrated that artificial oversulfation of DS increased its biological half-life after intravenous administration to rats.
Topics: Animals; Anticoagulants; Chromatography, High Pressure Liquid; Dermatan Sulfate; Disaccharides; Fluorescent Dyes; Guanidine; Guanidines; Half-Life; Magnetic Resonance Spectroscopy; Male; Rats; Rats, Wistar
PubMed: 8811915
DOI: 10.1006/abio.1996.0352 -
Methods in Molecular Biology (Clifton,... 2001
Topics: Animals; Chondroitin Lyases; Chondroitin Sulfates; Dermatan Sulfate; Disaccharides; Humans; Molecular Structure; Substrate Specificity
PubMed: 11450250
DOI: 10.1385/1-59259-209-0:363 -
Glycobiology Nov 2009Chondroitin/dermatan sulfate is a highly complex linear polysaccharide ubiquitously found in the extracellular matrix and at the cell surface. Several of its functions,...
Chondroitin/dermatan sulfate is a highly complex linear polysaccharide ubiquitously found in the extracellular matrix and at the cell surface. Several of its functions, such as binding to growth factors, are mediated by domains composed of alternating iduronic acid and 4-O-sulfated N-acetylgalactosamine residues, named 4-O-sulfated iduronic acid blocks. These domains are generated by the action of two DS-epimerases, which convert D-glucuronic acid into its epimer L-iduronic acid, in close connection with 4-O-sulfation. In this study, dermatan sulfate structure was evaluated after downregulating or increasing dermatan 4-O-sulfotransferase 1 (D4ST-1) expression. siRNA-mediated downregulation of D4ST-1 in primary human lung fibroblasts led to a drastic specific reduction of iduronic acid blocks. No change of epimerase activity was found, indicating that the influence of D4ST-1 on epimerization is not due to an altered expression level of the DS-epimerases. Analysis of the dermatan sulfate chains showed that D4ST-1 is essential for the biosynthesis of the disulfated structure iduronic acid-2-O-sulfate-N-acetylgalactosamine-4-O-sulfate, thus confirmed to be strictly connected with the iduronic acid blocks. Also the biologically important residue hexuronic acid-N-acetylgalactosamine-4,6-O-disulfate considerably decreased after D4ST-1 downregulation. In conclusion, D4ST-1 is a key enzyme and is indispensable in the formation of important functional domains in dermatan sulfate and cannot be compensated by other 4-O-sulfotransferases.
Topics: Dermatan Sulfate; Down-Regulation; Fibroblasts; Humans; Iduronic Acid; Lung; RNA, Small Interfering; Sulfotransferases
PubMed: 19661164
DOI: 10.1093/glycob/cwp110 -
Journal of Pharmaceutical and... Dec 2010Capillary electrophoresis (CE) was applied to the quantitation of dermatan sulfate (DS) and chondroitin sulfate (CS) as related substances in sodium heparin. The method... (Comparative Study)
Comparative Study
Capillary electrophoresis (CE) was applied to the quantitation of dermatan sulfate (DS) and chondroitin sulfate (CS) as related substances in sodium heparin. The method is based on the selective digestion of either CS and DS contained in the main drug heparin, by using chondroitinase ABC (specific for both DS and CS) and chondroitinase AC (specific for only CS). The unsaturated disaccharides released after exhaustive digestion, can be separated by CE using a 110mM phosphate buffer, pH 3.5 as the background electrolyte in a fused silica capillary (64.5cmx50mum i.d.) at 40 degrees C and -30kV. Since the level of each disaccharide released upon enzymatic digestion corresponds to its content in the native glycosaminoglycan, the amount of CS and DS was determined by proportion with the released disaccharides. In particular, DeltaUA-->GalNAc-4S Na(2) and DeltaUA-->GalNAc-6S Na(2) were selected for quantitation of CS and DS because of their significant response and short migration time (less than 7min).The method was validated for linearity, accuracy, precision and it showed to be able in detecting selectively, DS and CS at impurity level (LOD 0.01%, w/w). The proposed CE approach was finally applied to real samples. The results obtained were found in excellent correlation with those achieved by the analysis of the same samples using the official USP method based on high performance anion exchange chromatography (HPAEC) with pulsed amperometric detector.
Topics: Chemistry, Pharmaceutical; Chondroitin Sulfates; Dermatan Sulfate; Drug Contamination; Electrophoresis, Capillary; Heparin
PubMed: 20674212
DOI: 10.1016/j.jpba.2010.07.006 -
Journal of Biochemistry Feb 1975A simple chemical method for the determination of individual mucopolysaccharides in mixtures of dermatan sulfate and chondroitin sulfates by means of a single reagent...
A simple chemical method for the determination of individual mucopolysaccharides in mixtures of dermatan sulfate and chondroitin sulfates by means of a single reagent was established, utilizing the difference in reaction rates of these polysaccharides with orcinol. To each 1 ml of a sample mixture of standard dermatan sulfate and standard chondroitin sulfate (either 4- or 6-sulfate) was added 3 ml of orcinol reagent and the resulting solution was heated in a boiling-water bath. After 20 and 60 min reaction, absorbances at 660 nm were measured and the concentrations of individual mucopolysaccharides were calculated. High reproducibility was observed for the determination of dermatan sulfate in the presence of chondroitin sulfates. In addition, orcinol reaction for 90 min employing D-glucuronolactone as a standard appeared to be of practical value in the estimation of the uronic acid content of these mucopolysaccharides.
Topics: Carbazoles; Chondroitin; Dermatan Sulfate; Hydrolysis; Indicators and Reagents; Kinetics; Methods; Spectrophotometry; Time Factors; Uronic Acids
PubMed: 1126924
DOI: 10.1093/oxfordjournals.jbchem.a130738 -
Glycoconjugate Journal Aug 2013Chondroitin sulfate (CS) and dermatan sulfate (DS) interact with various extracellular molecules such as growth factors, cytokines/chemokines, neurotrophic factors,...
Chondroitin sulfate (CS) and dermatan sulfate (DS) interact with various extracellular molecules such as growth factors, cytokines/chemokines, neurotrophic factors, morphogens, and viral proteins, thereby playing roles in a variety of biological processes including cell adhesion, proliferation, tissue morphogenesis, neurite outgrowth, infections, and inflammation/leukocyte trafficking. CS/DS are modified with sulfate groups at C-2 of uronic acid residues as well as C-4 and/or C-6 of N-acetyl-D-galactosamine residues, yielding enormous structural diversity, which enables the binding with numerous proteins. We have demonstrated that highly sulfated CS-E from squid cartilage, for example, interacts with heparin-binding proteins including midkine, pleiotrophin, and fibroblast growth factors expressed in brain with high affinity (Kd values in the nM range). Here, we analyzed the binding of CS and DS, which have a relatively low degree of sulfation and have been widely used as a nutraceutical and a drug for osteoarthritis etc., with a number of heparin-binding neurotrophic factors/cytokines using surface plasmon resonance (SPR) and structurally characterized the CS/DS chains. SPR showed that relatively low sulfated CS-A, DS, and CS-C also bound with significant affinity to midkine, pleiotrophin, hepatocyte growth factor, monokine-induced by interferon-γ, and stromal cell derived factor-1β, although the binding was less intense than that with highly sulfated CS-D and CS-E. These findings suggest that even low sulfated CS and/or DS chains may contain binding domains, which include fine sugar sequences with specific sulfation patterns, and that sugar sequences, conformations and electrostatic potential are more important than the simple degree of sulfation represented by disaccharide composition.
Topics: Acetylgalactosamine; Animals; Cartilage; Chondroitin Sulfates; Cytokines; Decapodiformes; Dermatan Sulfate; Fibroblast Growth Factors; Kinetics; Protein Binding
PubMed: 23275130
DOI: 10.1007/s10719-012-9463-5 -
Organic & Biomolecular Chemistry Jun 2013Here, we present a novel approach for the chemical synthesis of chondroitin and dermatan sulfate oligosaccharides. A key point of this strategy is the preparation and...
Here, we present a novel approach for the chemical synthesis of chondroitin and dermatan sulfate oligosaccharides. A key point of this strategy is the preparation and use of an N-trifluoroacetyl galactosamine building block containing a 4,6-O-di-tert-butylsilylene group. Glycosylation reactions proceeded in good yields (74-91%) with our protecting group distribution. Using this approach, we have synthesized, for the first time, a chondroitin/dermatan sulfate-like tetrasaccharide that contains both types of uronic acids, D-glucuronic and L-iduronic acid. Moreover, we have employed a fluorescence polarization competition assay to evaluate the interactions between the synthesized oligosaccharides and FGF-2 (basic fibroblast growth factor). Our results show that this method, using standard instrumentation and minimal sample consumption, is a powerful tool for the rapid analysis of the glycosaminoglycan affinity for proteins in solution.
Topics: Chondroitin Sulfates; Dermatan Sulfate; Fibroblast Growth Factor 2; Fluorescence Polarization; Magnetic Resonance Spectroscopy; Molecular Structure; Protein Binding
PubMed: 23595496
DOI: 10.1039/c3ob40306h -
Pathophysiology of Haemostasis and... 2002Dermatan sulphate (DS) is a glycosaminoglycan which selectively catalyzes the inactivation of thrombin by Heparin Cofactor II without interacting with Antithrombin III.... (Review)
Review
Dermatan sulphate (DS) is a glycosaminoglycan which selectively catalyzes the inactivation of thrombin by Heparin Cofactor II without interacting with Antithrombin III. DS does not interact with other coagulation factors and, unlike heparin, is able to inactivate thrombin bound to fibrin or to the surface of an injured vessel. Efficacy and safety of DS have been validated in studies on thromboprophylaxis and on the anticoagulation for hemodialysis. Studies on thromboprophylaxis have been performed in "medical" patients as well as in general, orthopedic and oncological surgery. In this last setting, DS proved to be more efficacious than heparin, in the absence of excess bleeding. No statistically significant differences were observed between DS and heparin in hemodialysis. A low-molecular-weight DS,which shows a higher bioavailability after s.c. administration, has been tested in pilot studies on the treatment of venous thromboembolism with encouraging results. Two DS-containing compounds, sulodexide and, particularly, mesoglycan, have been clinically studied in a number of trials and found to be effective in the treatment of venous and arterial leg diseases.
Topics: Anticoagulants; Dermatan Sulfate; Humans; Thromboembolism
PubMed: 13679662
DOI: 10.1159/000073586 -
Journal of Neuropathology and... Aug 2014Widespread skeletal muscle degeneration and impaired regeneration lead to progressive muscle weakness and premature death in patients with Duchenne muscular dystrophy...
Widespread skeletal muscle degeneration and impaired regeneration lead to progressive muscle weakness and premature death in patients with Duchenne muscular dystrophy (DMD). Dystrophic muscles are progressively replaced by nonfunctional tissue because of exhaustion of muscle precursor cells and excessive accumulation of extracellular matrix (ECM). Sulfated glycosaminoglycans (GAGs) are components of the ECM and are increasingly implicated in the regulation of biologic processes, but their possible role in the progression of DMD pathology is not understood. In the present study, we performed immunohistochemical and biochemical analyses of endogenous GAGs in skeletal muscle biopsies of 10 DMD patients and 11 healthy individuals (controls). Immunostaining targeted to specific GAG species showed greater deposition of chondroitin sulfate (CS)/dermatan (DS) sulfate in DMD patient biopsies versus control biopsies. The selective accumulation of CS/DS in DMD biopsies was confirmed by biochemical quantification assay. In addition, high-performance liquid chromatography analysis demonstrated a modification of the sulfation pattern of CS/DS disaccharide units in DMD muscles. In conclusion, our data open up a new path of investigation and suggest that GAGs could represent a new and original therapeutic target for improving the success of gene or cell therapy for the treatment of muscular dystrophies.
Topics: Adolescent; Adult; Child; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Dermatan Sulfate; Disease Progression; Female; Glycosaminoglycans; Humans; Male; Muscle, Skeletal; Muscular Dystrophy, Duchenne; RNA, Messenger; Young Adult
PubMed: 25003237
DOI: 10.1097/NEN.0000000000000098