Isolation and partial characterization of a high molecular weight anionic glycoconjugate from transforming growth factor-beta treated bovine articular chondrocyte cultures.
A high molecular weight anionic glycoconjugate was isolated from the media of the transforming growth factor-beta treated chondrocyte cultures by anion-exchange chromatography on DEAE-Sephacel and Mono Q columns and was partially characterized. This high molecular weight anionic glycoconjugate was not detected in the non-treated (control) cultures. Characterization studies showed that the glycoconjugate is a non-reducible, non-collagenous glycoprotein containing O-linked, N-linked, and sialic acid substituted carbohydrate units. The isolated glycoconjugate stained "blue" with Stains All and migrated as a single band on sodium dodecyl sulfate gradient gels (2.5-10% acrylamide - diallyl tartardiamide) at an estimated molecular weight of 540 000. Amino acid and amino sugar analyses showed that it is rich in aspartic acid--asparagine, glutamic acid--glutamine, alanine, proline, and glycine, and contains galactosamine and glucoasmine. This transforming growth factor-beta inducible glycoprotein may be involved in cell differentiation and in the cartilage repair process. It may also be used as a marker to localize the biological activity of transforming growth factor-beta in articular cartilage.
Chan
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CK
Department of Medicine, Queen's University, Kingston, Canada.
Anastassiades
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D000596:Amino Acids / Q000032:analysis; D000818:Animals; D015415:Biomarkers; D002241:Carbohydrates / Q000032:analysis; D002358:Cartilage, Articular / Q000737:chemistry* / Q000187:drug effects; D002417:Cattle; D002478:Cells, Cultured; D002850:Chromatography, Gel; D017077:Culture Media, Conditioned / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification*; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D000596:Amino Acids; D015415:Biomarkers; D002241:Carbohydrates; D017077:Culture Media, Conditioned; D006023:Glycoproteins; D016212:Transforming Growth Factor beta; D019158:N-Acetylneuraminic Acid