Separation and characterization of two populations of aggregating proteoglycans from cartilage.
Intermediary gel immunoelectrophoresis was used to show that purified aggregating cartilage proteoglycans from 2-year-old steers contain two distinct populations of molecules and that only one of these is immunologically related to non-aggregating cartilage proteoglycans. The two types of aggregating proteoglycans were purified by density-gradient centrifugation in 3.5M-CsCl/4M-guanidinium chloride and separated by zonal rate centrifugation in sucrose gradients. The higher-buoyant-density faster-sedimenting proteoglycan represented 43% of the proteoglycans in the extract. It had a weight-average Mr of 3.5 X 10(6), did not contain a well-defined keratan sulphate-rich region, had a quantitatively dominant chondroitin sulphate-rich region and contained 5.9% protein and 23% hexosamine. The lower-buoyant-density, more slowly sedimenting, proteoglycan represented 15% of the proteoglycans in the extract. It had a weight-average Mr of 1.3 X 10(6), contained both the keratan sulphate-rich and the chondroitin sulphate-rich regions and contained 7.3% protein and 23% hexosamine. Each of the proteoglycan preparations showed only one band on agarose/polyacrylamide-gel electrophoresis. The larger proteoglycan had a lower mobility than the smaller. The distribution of chondroitin sulphate chains along the chondroitin sulphate-rich region was similar for the two types of proteoglycans. The somewhat larger chondroitin sulphate chains of the larger proteoglycan could not alone account for the larger size of the proteoglycan. Peptide patterns after trypsin digestion of the proteoglycans showed great similarities, although the presence of a few peptides not shared by both populations indicates that the core proteins are partially different.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002356:Cartilage / Q000032:analysis*; D002417:Cattle; D002501:Centrifugation, Zonal; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006595:Hexosamines / Q000032:analysis; D007122:Immunoelectrophoresis; D008027:Light; D046911:Macromolecular Substances; D008297:Male; D008970:Molecular Weight; D011509:Proteoglycans / Q000302:isolation & purification; D012542:Scattering, Radiation
D000596:Amino Acids; D006595:Hexosamines; D046911:Macromolecular Substances; D011509:Proteoglycans