Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
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The glycosaminoglycan (GAG) contents of hypertrophic scars, normal scars, and human skin from cadavers of matched ages were compared. Cellulose acetate electrophoresis, chondroitinase digestions, and reaction product and infrared analyses were used to characterize the component GAGs. DEAE-cellulose chromatography was used to separate hyaluronic acid (HA) and sulfated GAGs. Chondroitinase analysis was improved under these conditions. HA was determined enzymatically. Results showed an elevation of HA in hypertrophic scar. Dermatan sulfate was the major GAG in both scars and a slightly greater quantity was observed in the hypertrophic scar. Small amounts of chondroitin 4-sulfate and chondroitin 6-sulfate disaccharide constituents were also detected by the chondroitinase assay method and these were also elevated in hypertrophic scar. These results suggest that the GAGs of hypertrophic scar differ from normal scar and normal skin.
Exp Mol Pathol 1984 Feb
PMID:Glycosaminoglycans of normal and hypertrophic human scar. 669

A murine BALB/c IgG2a (lambda 3) myeloma immunoglobulin SAPC-15 with binding activity for negatively charged polysaccharides has been purified by affinity chromatography, and its interaction with heparin and various other polyanionic antigens has been studied. The antigen-binding activity has been demonstrated to reside in the Fab part of the immunoglobulin. The S15 myeloma protein in 0.05 M Tris buffer at pH 7.4 precipitated dextran sulfate, heparin, chondroitin sulfate A, B and C, hyaluronic acid, H. influenzae type b polysaccharide, calf thymus DNA, Klebsiella polysaccharide K63 and poly-L glutamic acid. Of these antigens only dextran sulfate was precipitated in 0.01 M phosphate buffered saline (0.15M), pH 7.4. The pepsin S15 Fab fragment did not precipitate with any of these antigens. The intrinsic tryptophanyl fluorescence of S15 was changed maximally by the addition of heparin, and the binding affinity of the immunoglobulin for this antigen was high (greater than 10(6) L/M). S15 may resemble antibody molecules that react with antigens under non-physiological conditions or in pathological conditions or in the external environment as in the lumen of the gut. All the above interactions of S15 with antigens persisted in 0.05 M Tris buffer made physiologically isotonic by the addition of sucrose, and S15 could thus be used to identify these antigens on cell surfaces.
Mol Immunol 1982 Jul
PMID:The interaction of mouse myeloma immunoglobulin S15 with negatively charged polysaccharide antigens. 681 62

C-reactive protein (CRP) is a trace serum protein which elevates up to 1000-fold in concentration in association with inflammation and tissue necrosis. CRP binds with phosphocholine and phosphate esters; initiates reactions of agglutination, opsonization and complement consumption; and precipitates with protamine and synthetic polymers of lysine and arginine, and these reactivities are modulated by calcium and phosphocholine. We now report on the interactions of heparin with these polycations in the absence and presence of CRP, which show marked similarities to reactions between antigen and antibody. Heparin optimally precipitated with the polycations over a narrow range of reactant ratios, peaking at slight anion charge excess. The complexes formed did not dissociate in excess heparin: however, complex formation was significantly depressed when heparin was added in a single dose in excess of the amount required for optimal precipitation. Calcium did not affect the precipitation of heparin with polycation, and heparin did not precipitate with CRP. However, heparin did induce a rapid and efficient dissociation of CRP-polycation precipitates preformed at equivalence, with total release of CRP. Small amounts of heparin augmented precipitation under conditions of polycation excess of CRP, but as heparin levels were increased to amounts needed to reach equivalence with polycation, CRP was resolubilized in favor of the preferred heparin-polycation complexes. Chondroitin sulfate was similar to heparin in its effects upon the CRP-poly-L-arginine (PLA) interaction, while hyaluronic acid enhanced CRP-PLA precipitation at all concentrations tested and DNA had neither augmenting nor solubilizing effects. These data indicate that CRP-polycation interactions are significantly and selectively influenced in the presence of small amounts of heparin and certain other polyanions. This modulatory reactivity may be of importance in the biological reactivities of CRP during episodes of acute inflammation.
Mol Immunol 1983 May
PMID:Influence of heparin on interactions between C-reactive protein and polycations. 687 43

Vanadate (50 microM) increases the incorporation of 3H from D-[1,6-3H]glucosamine into hyaluronic acid (HA) in serum stimulated Swiss 3T3 fibroblasts. This increase is not a result of increased HA synthesis, but is due to a significant increase in the specific activity of the cellular UDP-N-acetyl-glucosamine. When the 3H incorporation is corrected for the change in precursor specific activity, vanadate is shown to inhibit the synthesis of HA, a result that is confirmed by measuring the HA with a competitive binding assay. The inhibition of HA synthesis by vanadate is also shown to be dependent on the media used in the experiment, with a substantial increase in the inhibition seen when the media contains the pH indicator, phenol red. The HA synthetase activity of isolated membranes is not inhibited by vanadate at concentrations up to 500 microM.
Biochem Mol Biol Int 1993 Nov
PMID:Vanadate inhibition of hyaluronic acid synthesis in Swiss 3T3 fibroblasts. 750 61

Hyaluronic acid, a major component of the extracellular matrix, plays an important role in the regulation of different cellular processes, e.g., locomotion, cell-cell interaction during morphogenesis, and differentiation. Distribution of hyaluronic acid with respect to the role of sperm hyaluronidase in sperm penetration and gamete interaction is well established. In order to elucidate this mechanism, in our current study we have identified and demonstrated, for the first time, the presence of a 68-kDa cell surface hyaluronic acid binding glycoprotein (HABP) in spermatozoa of different species (rat, mice, bull, and human) by immunoblot analysis and indirect immunofluorescence using the polyclonal antibodies raised against purified HABP. Furthermore, we were able to demonstrate a differential distribution of 68-kDa HA binding protein on the sperm head, midpiece, and tail of different species. To identify its role in sperm function, we observed its declining pattern during epididymal maturation and also the inhibition of sperm-oolemmal adherence by pretreatment of the sperms with anti-HABP antibodies. We have further observed its in vivo phosphorylation in motile spermatozoa. All our data clearly indicate that sperm hyaluronan binding protein may have a specific role in sperm maturation, motility, and fertilization processes.
Mol Reprod Dev 1994 May
PMID:Evidence for presence of hyaluronan binding protein on spermatozoa and its possible involvement in sperm function. 751 32

An improved assay for hyaluronic acid (HA) synthetase is described that is suitable for rapid processing of large numbers of samples. High background levels of unincorporated radioactivity are removed by passage of the reaction through a Sephadex G-50 spin column. The labeled HA product is then precipitated onto glass fiber filters with cetylpyridinium chloride. Apparent Km values for HA synthetase from Swiss 3T3 fibroblasts are 10.8 and 58.4 microM for UDP-glucuronic acid and UDP-N-acetylglucosamine, respectively. HA synthetase activity of quiescent cells is 4.5% of that found in actively growing cells and is stimulated in response to 10% calf serum. There is a greater than 10-fold increase in HA synthetase activity when cells are harvested with hyaluronidase as compared with trypsin.
Biochem Mol Biol Int 1995 Apr
PMID:A filter paper assay for hyaluronic acid synthetase: application to the enzyme from Swiss 3T3 fibroblasts. 754 31

Significant changes in the synthesis of glycosaminoglycans occur during the transformation of stromal cells of the endometrium into decidual cells which takes place during the initial stages of pregnancy in mice. Hyaluronic acid, which is practically absent in the endometrium of virgin mice, increases dramatically on the fifth day of pregnancy, reaching its maximal concentration on day 6 followed by a 50% decrease on day 7. Changes in hyaluronic acid concentration also occur in pseudopregnant mice indicating that they are not related to the presence of the embryo in the uterus. The absolute concentration of the sulfated glycosaminoglycans, e.g., heparan sulfate, dermatan sulfate and chondroitin sulfate in the decidua did not change significantly. There was, however, a striking decrease of their biosynthesis in pregnant and pseudopregnant mice when compared to virgin mice, as shown by the use of radioactive inorganic sulfate as a precursor for the study of in vivo synthesis. A radioautographical analysis confirmed that the highest incorporation of radioactive sulfate was observed in virgin endometria when compared to pregnant ones. These studies also have shown a characteristic pattern of labeling in different regions of the endometrium that repeats itself during the different days of pregnancy.
Cell Mol Biol (Noisy-le-grand) 1995 Feb
PMID:Biosynthesis of glycosaminoglycans in the endometrium during the initial stages of pregnancy of the mouse. 777 41

The important components of mucopolysaccharides and collagen have been analyzed in tissues of control and carcinoma of uterine cervix. Among these components hyaluronic acid and chondroitin sulphate levels were found to be increased, whereas decreased level of collagen was observed in uterine cervical carcinoma. Serum cathepsin B, D and acid and alkaline phosphatases have also been analyzed in controls and carcinoma patients before and after treatments. The activities of these enzymes have been found to increase prominently in advanced stages. Among these enzymes cathepsin B and alkaline phosphatase have exhibited remarkable increase in activity in uterine cervical carcinoma. Different modes of treatment exerted reversion of the elevated activities of these enzymes. However, combined therapy type II (radiation combined with cisplatin and cyclophosphomide) seems to be more effective in reverting the activities of these enzymes to normal levels.
Mol Cell Biochem 1995 Mar 09
PMID:Extracellular matrix components and proteolytic enzymes in uterine cervical carcinoma. 779 43

The parallel fiber "en passant" synaptic endings of mouse cerebellar molecular layer have shown by means of transmission electron microscopy, the presence of an electron dense extravesicular material in samples perfused with Alcian blue. This alcianophilic material was digested in cerebellar tissue previously treated with testicular hyaluronidase, suggesting the presence of hyaluronic acid or chondroitin 4- or 6-sulphate. Freeze-fractured Rhesus monkey cerebellar cortex prepared for conventional scanning electron microscopy also revealed the presence in fractured synaptic varicosities of parallel fibers of a high mass density material, in which the synaptic vesicles are embedded. Examination of cryofractured primate cerebellar cortex coated with thin chromium films, 1-2 nm thick, in the high resolution field emission scanning electron microscope showed the SE-I topographic contrast of an extravesicular material deposited in axoplasmic matrix of fractured parallel synaptic endings. The precise localization of this material corresponds to that observed in transmission electron microscopy and conventional freeze-fracture scanning electron microscopy. These electron microscopic findings tend to agree with the omnipresence in several vertebrates of a presynaptic axoplasmic material, which seems to be proteoglycan in nature.
Cell Mol Biol (Noisy-le-grand) 1994 Sep
PMID:Proteoglycan ultracytochemistry and conventional and high resolution scanning electron microscopy of vertebrate cerebellar parallel fiber presynaptic endings. 781 87

A highly sensitive method for detection of the carbohydrate-binding activity of proteins is described. The method is based on interactions of carbohydrate-binding proteins, immobilized on a solid phase, with an enzyme-labeled soluble polysaccharide (peroxidase conjugated glycosaminoglycans-heparin, chondroitin sulfate or hyaluronic acid. Binding capacity was measured spectrofotometrically after enzymatic reaction with chromogenic substrate. The reliability of the assay was tested by use of two heparin-binding proteins-i) fibronectin (soluble) and ii) heparin-binding protein purified from the human brain (water-insoluble). Binding of heparin was dependent on metal ions, detergents and urea. The assay is believed to be applicable for the identification and characterization of a variety of carbohydrate(glycosaminoglycan)-binding proteins, especially, when traditional methods can not be applied (e.g., when proteins are water-insoluble).
Biochem Mol Biol Int 1994 Sep
PMID:A novel method for evaluation of carbohydrate-binding activity: enzyme-linked carbohydrate-binding assay (ELCBA). 784 36


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