Structure of proteoheparan sulfates from fibroblasts. Confluent and proliferating fibroblasts produce at least three types of proteoheparan sulfates with functionally different core proteins.
[3H]Leucine- and [35S]sulfate-labeled proteoheparan sulfates were isolated from postconfluent or proliferating cultures of human skin fibroblasts. Cell layers were solubilized by Triton X-100, and transferrin-binding macromolecules were isolated by affinity chromatography. Proteoglycans with no affinity for transferrin were purified by using ion-exchange and gel permeation chromatography. Postconfluent cells synthesize a proteoheparan sulfate of Mr 350,000 (as determined by gel permeation chromatography) which has affinity for transferrin as well as for octyl-Sepharose. Its core protein (Mr 180,000) consists of two disulfide-bonded polypeptides of Mr 90,000. This species was not detected in cultures of proliferating cells. Proliferating and confluent cells also synthesize other forms of proteoheparan sulfates (Mr 200,000-400,000) which have no affinity for transferrin. However, most of them have affinity for octyl-Sepharose. The core protein of proteoheparan sulfates made by proliferating cells has Mr 50,000. A smaller form (Mr 250,000) of this proteoglycan was solubilized by Triton X-100, whereas a larger form (Mr 400,000) remained associated with the pericellular matrix. A third type of proteoheparan sulfate (Mr 200,000) without affinity for transferrin nor octyl-Sepharose was associated with postconfluent cell layers but not with proliferating ones. Its core protein has Mr 35,000. Heparan sulfate oligosaccharides (Mr 6,000 or higher) were found in proliferating cells but not in postconfluent ones.
D002455:Cell Division; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis*; D005347:Fibroblasts / Q000032:analysis*; D006025:Glycosaminoglycans / Q000032:analysis*; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000032:analysis*; D006801:Humans; D007930:Leucine / Q000378:metabolism; D008970:Molecular Weight; D017830:Octoxynol; D011092:Polyethylene Glycols; D011509:Proteoglycans / Q000032:analysis*; D011956:Receptors, Cell Surface / Q000302:isolation & purification; D011990:Receptors, Transferrin; D013431:Sulfates / Q000378:metabolism
D011508:Chondroitin Sulfate Proteoglycans; D006025:Glycosaminoglycans; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D007930:Leucine; D017830:Octoxynol; D011092:Polyethylene Glycols; D011509:Proteoglycans; D011956:Receptors, Cell Surface; D011990:Receptors, Transferrin; D013431:Sulfates