Bovine aorta contains at least two related forms of heparan sulphate proteoglycan.
1. The proteoglycan peak from anion exchange chromatography of an extract of bovine aorta was digested with chondroitinase ABC. The residual heparan sulphate proteoglycans were further purified by chromatography on Sepharose CL4B and DEAE-Sephacel to yield two species, of high and low charge density. 2. Higher molecular weight material had a higher proportion of high charge density proteoglycan, while the lower molecular weight species had a higher proportion of low charge density heparan sulphate proteoglycan. 3. The two species shared epitopes as they both reacted with an antibody to heparan sulphate proteoglycan from bovine glomerular basement membrane. 4. On electron microscopy, both high and low charge density proteoglycans were visualized as 'tadpole-like' molecules, which showed a tendency to aggregate via their globular heads. 5. Bovine aortic smooth muscle cells were cultured in the presence of [35S]sulphate and [3H]glucosamine. Proteoglycans were isolated from medium and cell layer extract by the methods outlined above. 6. The major HSPG species isolated from medium were significantly larger than those from cell layer and displayed substantial heterogeneity in both size of HS chain after papain digestion and size of protein core after heparitinase digestion. 7. The major cell layer species yielded two HS species of widely differing mol. wt after papain digestion, and a very small protein core after heparitinase digestion. Therefore cell layer-associated HSPGs show a good deal more homogeneity than those found in the medium. 8. Further ion-exchange chromatography after digestion with chondroitinase ABC revealed HSPG species of lower charge density, possibly derived from a hybrid chondroitin sulphate-dermatan sulphate proteoglycan (CS/DSPG) after removal of the CS/DS chains.
Lindsay
C D
CD
Department of Biological Sciences, Medical School University of Manchester, U.K.
Anderson
J C
JC
D000818:Animals; D001013:Aorta, Thoracic / Q000737:chemistry*; D002417:Cattle; D002808:Chondroitin Lyases; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004797:Enzyme-Linked Immunosorbent Assay; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000737:chemistry*; D008854:Microscopy, Electron; D011509:Proteoglycans / Q000737:chemistry*
D002808:Chondroitin Lyases; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D011509:Proteoglycans