Analysis of sulfur-35-labeled proteoglycans indicated that cholesterol-enriched pigeon peritoneal macrophages synthesized 42% more 35S-labeled proteoglycan when compared with control macrophages during a 24-hour incubation. Proteoglycan turnover was subsequently studied in radiolabeled macrophage cultures after a 1-, 3-, 6-, 12-, or 24-hour chase with fresh media. During the chase, intracellula...

Sonic hedgehog promotes proliferation of developing cerebellar granule cells. As sonic hedgehog is expressed in the cerebellum throughout life it is not clear why proliferation occurs only in the early postnatal period and only in the external granule cell layer. We asked whether heparan sulfate proteoglycans might regulate sonic hedgehog-induced proliferation and thereby contribute to the spec...

To study how cells regulate the composition of glycosaminoglycan chains on proteoglycans, we have examined the assembly of chains on chimeric proteoglycans containing segments of betaglycan (transforming growth factor-beta Type III receptors) fused to protein A. Transient expression of the chimeras in Chinese hamster ovary cells revealed that only two glycosaminoglycan attachment sites exist. O...

Kidneys were perfused with [35S]sulfate at 4 h in vitro to radiolabel sulfated proteoglycans. Glomeruli were isolated from the labeled kidneys, and purified fractions of glomerular basement membrane (GBM) were prepared therefrom. Proteoglycans were extracted from GBM fractions by use of 4 M guanidine-HCl at 4 degrees C in the presence of protease inhibitors. The efficiency of extraction was app...

Proteoglycans were extracted from bovine lung gas exchange tissue, pleura, and bronchioles with 4.0 M guanidinium chloride at 5 degrees C in the presence of protease inhibitors. Preliminary purification of the proteoglycans was achieved by an initial CsCl isopycnic centrifugation (rho 0 = 1.33) and through precipitation with cetylpyridinium chloride in 0.5 M KCl. Further purification and fracti...

Proteoglycans located in basement membranes, the nanostructures underling epithelial and endothelial layers, are unique in several respects. They are usually large, elongated molecules with a collage of domains that share structural and functional homology with numerous extracellular matrix proteins, growth factors and surface receptors. They mainly carry heparan sulfate side chains and these c...

The method of affinity coelectrophoresis was used to study the binding of nine representative glycosaminoglycan (GAG)-binding proteins, all thought to play roles in nervous system development, to GAGs and proteoglycans isolated from developing rat brain. Binding to heparin and non-neural heparan and chondroitin sulfates was also measured. All nine proteins-laminin-1, fibronectin, thrombospondin...

The distribution of proteoglycans in the trabecular tissue of human eyes was evaluated histochemically, using Cupromeronic Blue in combination with a series of enzyme digestions and nitrous acid treatment. Within the extracellular matrix of the trabecular meshwork, many Cupromeronic Blue-positive filaments were observed in association with collagen fibrils, basal lamina, a basal lamina-like mat...

We used antibodies raised against both a heparan sulfate proteoglycan purified from a mouse sarcoma and a chondroitin sulfate proteoglycan purified from a rat yolk sac carcinoma to study the appearance and distribution of proteoglycans in cultured cells. Normal rat kidney cells displayed a fibrillar network of immunoreactive material at the cell surface when stained with antibodies to heparan s...