title: Isolation and identification of the major heparan sulfate proteoglycans in the developing bovine rib growth plate.
abstract: Heparan sulfate proteoglycans are thought to mediate the action of growth factors. The heparan sulfate-containing proteoglycans in extracts of the bovine fetal rib growth plate were detected using the monoclonal antibody 3G10, which recognizes a neoepitope generated by heparitinase digestion (David, G., Bai, X. M., Van der Schueren, B., Cassiman, J. J., and Van den Berghe, H. (1992) J. Cell Biol. 119, 961-975). The heparan sulfate proteoglycans that react with this antibody were identified using antisera to known proteoglycans; purified using CsCl density gradient centrifugation, molecular sieve, and ion exchange chromatography; and then characterized. The major heparan sulfate proteoglycans in the growth plate had core proteins of 200 kDa and larger and were identified as perlecan and aggrecan. These two heparan sulfate proteoglycans could be effectively separated from each other by CsCl density gradient centrifugation alone. Perlecan contained 25% heparan sulfate and 75% chondroitin sulfate. The heparan sulfate chains on growth plate perlecan were considerably smaller than the chondroitin sulfate chains, and the heparan sulfate disaccharide content was different than that found for heparan sulfate from either kidney, tumor tissue, or growth plate aggrecan. Aggrecan contained only 0.1% heparan sulfate, which was localized to the CS-1 domain of aggrecan. These results indicate that perlecan and aggrecan would be the principal candidate proteoglycans involved in the action of heparan sulfate-binding proteins in the developing growth plate.
authors.affiliation: Center for Research in Skeletal Development and Pediatric Orthopedics, Shriners Hospital for Children, Tampa, Florida 33612, USA.
authors.forename: Prasanthi
authors.initials: P
authors.lastname: Govindraj
authors.forename: Leigh
authors.initials: L
authors.lastname: West
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Koob
authors.forename: Peter
authors.initials: P
authors.lastname: Neame
authors.forename: Kurt
authors.initials: K
authors.lastname: Doege
authors.forename: John R
authors.initials: JR
authors.lastname: Hassell
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002586:Cesium / Q000494:pharmacology; D002712:Chlorides / Q000494:pharmacology; D002809:Chondroitin Sulfates / Q000737:chemistry; D002852:Chromatography, Ion Exchange; D000939:Epitopes; D019812:Heparan Sulfate Proteoglycans / Q000737:chemistry*; D007668:Kidney / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011509:Proteoglycans / Q000378:metabolism; D012272:Ribs / Q000196:embryology*; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology
chemical_list: D002712:Chlorides; D000939:Epitopes; D019812:Heparan Sulfate Proteoglycans; D010455:Peptides; D011509:Proteoglycans; C071927:perlecan; D002586:Cesium; D002809:Chondroitin Sulfates; D014357:Trypsin; C028019:cesium chloride