PMID: 2499259Jun 1, 1989Paper

Studies on the hyaluronate binding properties of newly synthesized proteoglycans purified from articular chondrocyte cultures

Archives of Biochemistry and Biophysics
J D Sandy, Anna Plaas

Abstract

Primary cultures of rabbit articular chondrocytes have been maintained for 10 days and labeled with [35S]sulfate, [3H]leucine, and [35S]cysteine in pulse-chase protocols to study the structure and hyaluronate binding properties of newly synthesized proteoglycan monomers. Radiolabeled monomers were purified from medium and cell-layer fractions by dissociative CsCl gradient centrifugation with bovine carrier monomer, and analyzed for hyaluronate binding affinity on Sepharose CL-2B in 0.5 M Na acetate, 0.1% Triton X-100, pH 6.8. Detergent was necessary to prevent self-association of newly synthesized monomers during chromatography. Monomers secreted during a 30-min pulse labeling with [35S]sulfate had a low affinity relative to carrier. Those molecules released into the medium during the first 12 h of chase (about 40% of the total) remained in the low affinity form whereas those retained by the cell layer rapidly acquired high affinity. In cultures where more than 90% of the preformed cell-layer proteoglycan was removed by hyaluronidase digestion before radiolabeling the newly synthesized low affinity monomers also rapidly acquired high affinity if retained in the cell layer. Cultures labeled with amino acid precursors were used t...Continue Reading

References

Jan 1, 1979·The Biochemical Journal·T E Hardingham
Aug 1, 1987·The Biochemical Journal·M PaulssonJ Engel
May 15, 1985·The Biochemical Journal·F BonnetT E Hardingham
Dec 1, 1985·The Biochemical Journal·J D Sandy, A H Plaas
Jan 1, 1982·Connective Tissue Research·R W FarndaleA J Barrett
Jul 1, 1980·Analytical Biochemistry·C B CaputoV C Hascall
Dec 1, 1981·Biochemical Society Transactions·I A Nieduszynski, J K Sheehan

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Citations

Mar 1, 1992·Biopolymers·R M Peitzsch, W F Reed
Mar 1, 1994·Matrix Biology : Journal of the International Society for Matrix Biology·K G VogelJ R Robbins
Feb 1, 1993·The Journal of Cell Biology·C B Knudson
May 15, 1993·Experientia·N S Fedarko
Jan 1, 1996·Connective Tissue Research·M P Maleski, C B Knudson
Jan 13, 2006·Osteoarthritis and Cartilage·M C StewartJ D Sandy
Aug 15, 1992·Archives of Biochemistry and Biophysics·C FlanneryJ Sandy
Mar 14, 1998·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·A C ChenR L Sah
Jul 1, 1990·Biological Chemistry Hoppe-Seyler·H S BrandJ K van der Korst
Sep 1, 1996·The American Journal of Physiology·H J HäuselmannE J Thonar
Jun 1, 1992·Clinical Biochemistry·K E Kuettner

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