Heparan sulfates of cultured cells. II. Acid-soluble and -precipitable species of different cell lines

Kraemer, Paul M.

doi:10.1021/bi00784a027

Cited by 206 publications

(51 citation statements)

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“…1 cells are described in Table 2. macromolecule was limited to those able to bind electron-dense stain (ruthenium red) (11) or to purely physical observations (4). Similarly, the presence of heparan sulfate in several tumor cell lines has been reported, but direct comparison with control cells has not been reported (12,13).…”

Section: And Discussionmentioning

confidence: 99%

Production of Mucopolysaccharides by Normal and Transformed Cells

Satoh

Duff

Rapp

et al. 1973

Proc. Natl. Acad. Sci. U.S.A.

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The rate of hyaluronic acid and sulfated mucopolysaccharide production was measured for hamster embryo fibroblasts and for general oncogenic lines derived by virus transformation. A striking increase in both the rate of hyaluronic acid synthesis and the amount of cellassociated polymer was observed after transformation by herpes simplex type-2 or SV40 virus. Although no corresponding change was observed for the sulfated polysaccharides, the proportion of heparan sulfate increased significantly after transformation.Recent reports from several laboratories have described effects on cell coat properties associated with virus transformation (1-3). In general, the morphologic observations have been attributed to changes in the content of those acid polysaccharides identified with the extracellular matrix-hyaluronic acid and a group of sulfated saccharides. Although differences in the rate of synthesis of cell coat have been observed for transformed cells (4), as a rule the polysaccharides involved have not been characterized.The present study describes the production and characterization of mucopolysaccharides by normal and virus-transformed fibroblasts from hamster embryo under cell culture conditions. An attempt was made to differentiate material adhering to cell surfaces from that released to the culture medium, although only an operational distinction was used.In most of the experiments reported, [3H]glucosamine was used as a radioactive precursor for both nonsulfated (hyaluronic acid) and sulfated (chondroitin sulfates, heparan sulfate, etc.) polysaccharides.The identification of the polysaccharides was based on elution behavior from cetylpyridinium chloride precipitates (5), high-voltage electrophoresis, enzyme susceptibilities, and characterization of the amino sugar component after acid hydrolysis. Table 1 illustrates the results obtained for several polysaccharides under consideration. MATERIALS AND METHODSSyrian hamster-embryo fibroblasts cells (inbred LSH strain, Lakeview Hamster Colony, New Field, N.J.) were prepared by dispersion of 13-day-old embryos with 0.25% trypsin. The dispersed cells were grown in medium 199 supplemented with 10% tryptose phosphate broth, 10% fetal-bovine serum, and 0.075% NaHCO3. Primary African green monkey kidney cells were also prepared by dispersion with 0.25% trypsin, and grown in Eagle's medium (BME, Flow Laboratories Inc.) with 10% fetal-calf serum and 0.075% NaHCO3.* To whom reprints requests should be addressed.The H-50 cell line was developed from a Syrian hamster tumor that was originally induced by SV40 (6). H-50 cells contain the SV40 T and transplantation-rejection antigens, and induce tumors when reinjected into weanling Syrian hamsters. They do not produce infectious virus. The cells were maintained in the identical manner as primary hamster embryo fibroblasts. The SV40-transformed BSC-1 cells are the results of the transformation of a stable line of African green monkey kidney cells (BSC-1) by SV40 (7). These cells synthesize the SV40-specific tumor antig...

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Section: And Discussionmentioning

confidence: 99%

Production of Mucopolysaccharides by Normal and Transformed Cells

Satoh

Duff

Rapp

et al. 1973

Proc. Natl. Acad. Sci. U.S.A.

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“…Many other cells also possess mucopolysaccharides on their surfaces (Kojima and Yamagata, 1971;Kraemer, 1971). Furthermore, virally transformed cells have varying amounts of extracellular mucopolysaccharides (Goggins et al, 1972;Satoh et al, 1973).…”

Section: Addition Of Chondroitin Sulfatementioning

confidence: 99%

Effect of Extracellular Chondroitin Sulfate on Cultured Chondrocytes

Huang

1974

The Journal of Cell Biology

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“…As far as we are aware, this represents the first description of sulfated GAG in the GBM, and, in addition, the first description of heparan sulfate in any basement membrane. Heparan sulfate has been found, however, in association with the cell surfaces of various different cell types (34), including vascular endothelia (35). The question arises as to why GAG have not been detected heretofore in the GBM either by biochemical assays (8)(9)(10) or by histochemical staining procedures (4-7, 11-12).…”

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confidence: 99%

Presence of heparan sulfate in the glomerular basement membrane

Kanwar

Farquhar

1979

Proc. Natl. Acad. Sci. U.S.A.

495

191

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The glomerular basement membrane was subjected to digestion with specific enzymes to determine the chemical nature (sialoglycoproteins, collagenous peptides, or glycosaminoglycans) of the anionic sites previously demonstrated in the laminae rarae. Enzyme digestion was carried out both in situ and in vitro. Kidneys were perfused in situ with enzyme solutions followed by perfusion with fixative containing the cationic dye, ruthenium red, to detect the anionic sites. Glomerular basement membranes were isolated by detergent treatment of glomeruli and incubated with enzyme solutions, followed by incubation with cationized ferritin (pI 7.3-7.5 Physiologic studies have established that the glomerular capillaries function as a size-and charge-selective barrier in the production of the glomerular filtrate and in the retention of plasma proteins in the circulation (1-3). By inference it has been assumed (1-3) that charge selectivity is due to the presence in the glomerulus of fixed negative charges, but their precise nature and location in the capillary wall have not been established. Since macromolecular compounds carrying sialic acid residues have been-up to recently-the only known polyanions present in the glomerulus, they have been considered the most likely candidates responsible for the charge barrier (1, 2, 4-7). Sialyl residues have been found in all three layers of the capillary wall. They have been detected in isolated glomerular basement membranes (GBM) (8-10) and in association with the cell membranes of both the endothelium (11-12) and epithelium (4-7, 11), where they are part of the sialic acid-rich, cell-surface coats. A colloidal iron-stainable, cell-coat material is particularly concentrated on the epithelium. Accordingly, it has been repeatedly suggested (1, 2, 4-7) that this "epithelial polyanion" is responsible for establishing the charge-selective properties of the glomerular filter, and, concomitantly, that its loss leads to proteinuria since loss of epithelial cell-coat staining occurs in several renal diseases (1, 2, 5). Materials. Ruthenium red was purchased from Ventron Corp. (Danvers, MA); horse-spleen ferritin (2X crystallized, cadmium free) from Calbiochem; and chondroitin sulfates (types A, B, and C) and heparin (grade I) from Sigma. Chondroitinase ABC (Proteus vulgaris) was obtained from Miles; testicular hyaluronidase from Sigma; leech hyaluronidase from Biotrics, Inc. (Boston, MA); neuraminidase (Clostridium perfringens, NEUA) from Worthington; collagenase form III (Cl. histolyticum) from Advance Biofactures (Lynbrook, NY); and Pronase (Streptomyces griseus, grade B) from Calbiochem. Crude heparinase and purified heparitinase (Flavobacterium heparinium) were the generous gift of Alfred Linker (17).* Neuraminidase and heparitinase were tested by the Azocoll assay (18) and were free of proteolytic activity. Cationized ferritin was prepared as described previously (16).Perfusion Experiments. The left kidney was exposed and perfused as described (16). Initially, the kidney was flush...

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Heparan sulfates of cultured cells. II. Acid-soluble and -precipitable species of different cell lines

Cited by 206 publications

References 3 publications

Production of Mucopolysaccharides by Normal and Transformed Cells

Production of Mucopolysaccharides by Normal and Transformed Cells

Effect of Extracellular Chondroitin Sulfate on Cultured Chondrocytes

Presence of heparan sulfate in the glomerular basement membrane

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