title: In situ cross-linking of cartilage proteoglycans.
abstract: Although the in vitro interactions between purified cartilage matrix components have been studied extensively, little is known about these interactions in situ. In this study, cartilage was treated with a cross-linking reagent with a span of 1.2 nm between its reactive terminal groups in order to preserve the native relationships between closely associated matrix components throughout extraction, purification, and preparation for electron microscopy. After in situ cross-linking, electron microscopy and gel chromatography revealed that about one-half of the guanidine hydrochloride extractable proteoglycans were polymeric, usually with two to five proteoglycan subunits in each polymer. Cross-linking consistently involved the thin segments of the proteoglycan subunits. Some of the proteoglycan polymers were capable of binding hyaluronic acid and were parts of aggregates under associative conditions. SDS-polyacrylamide gel electrophoresis revealed that link proteins were present within the polymers, and studies in which purified proteoglycans were cross-linked in vitro confirmed that the link proteins increased the proportion of polymeric proteoglycans. These findings suggest that individual proteoglycans within cartilage have intimate associations with other proteoglycans that are mediated by link proteins.
authors.affiliation: Department of Orthopaedic Surgery and Rehabilitation, University of Chicago, Illinois.
authors.forename: F M
authors.initials: FM
authors.lastname: Phillips
authors.forename: L A
authors.initials: LA
authors.lastname: Pottenger
authors.forename: R V
authors.initials: RV
authors.lastname: Hay
mesh_terms: D000818:Animals; D002356:Cartilage / Q000032:analysis* / Q000187:drug effects; D002417:Cattle; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D008239:Lysine / Q000378:metabolism; D008854:Microscopy, Electron; D011509:Proteoglycans / Q000032:analysis* / Q000378:metabolism
chemical_list: D003432:Cross-Linking Reagents; D011509:Proteoglycans; D008239:Lysine