The cell surface proteoglycan from mouse mammary epithelial cells bears chondroitin sulfate and heparan sulfate glycosaminoglycans.
The cell surface proteoglycan fraction isolated by mild trypsin treatment of NMuMG mouse mammary epithelial cells contains largely heparan sulfate, but also 15-24% chondroitin sulfate glycosaminoglycans. We conclude that this fraction contains a unique hybrid proteoglycan bearing both heparan sulfate and chondroitin sulfate glycosaminoglycans because (i) the proteoglycan behaves as a single species by sizing, ion exchange and collagen affinity chromatography, and by isopycnic centrifugation, even in the presence of 8 M urea or 4 M guanidine hydrochloride, (ii) the behavior of the chondroitin sulfate in these separation techniques is affected by heparan sulfate-specific probes and vice versa, and (iii) proteoglycan core protein bearing both heparan sulfate and chondroitin sulfate is recognized by a single monoclonal antibody. Removal of both types of glycosaminoglycan reduces the proteoglycan to a core protein of approximately 53 kDa. The proteoglycan fraction is heterogeneous in size, largely due to a variable number and/or length of the glycosaminoglycan chains. We estimate that one or two chondroitin sulfate chains (modal Mr of 17,000) exist on the proteoglycan for every four heparan sulfate chains (modal Mr of 36,000). Synthesis of these chains is reportedly initiated on an identical trisaccharide that links the chains to the same amino acid residues on the core protein. Therefore, some regulatory information, perhaps residing in the amino acid sequence of the core protein, must determine the type of chain synthesized at any given linkage site. Post-translational addition of these glycosaminoglycans to the protein may provide information affecting its ultimate localization. It is likely that the protein is directed to specific sites on the cell surface because of the ability of the glycosaminoglycans to recognize and bind extracellular components.
Rapraeger
A
A
Jalkanen
M
M
Endo
E
E
Koda
J
J
Bernfield
M
M
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis; D002807:Chondroitin / Q000031:analogs & derivatives*; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis*; D002809:Chondroitin Sulfates / Q000032:analysis*; D003094:Collagen / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004848:Epithelium / Q000032:analysis; D006025:Glycosaminoglycans / Q000032:analysis*; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000032:analysis*; D008321:Mammary Glands, Animal / Q000032:analysis* / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D009608:Nitrous Acid / Q000494:pharmacology; D011509:Proteoglycans / Q000032:analysis* / Q000302:isolation & purification* / Q000378:metabolism
D002807:Chondroitin; D011508:Chondroitin Sulfate Proteoglycans; D002809:Chondroitin Sulfates; D003094:Collagen; D004229:Dithiothreitol; D006025:Glycosaminoglycans; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D009608:Nitrous Acid; D011509:Proteoglycans