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[18] Biosynthesis of cartilage proteoglycan and link protein

1987; Academic Press; Linguagem: Inglês

10.1016/0076-6879(87)44187-6

1557-7988

James H. Kimura, Tamayuki Shinomura, Eugene J.‐M.A. Thonar,

Osteoarthritis Treatment and Mechanisms

This chapter describe three procedures which have proved useful in the isolation and characterization of newly synthesized proteoglycan and link protein. The first deals with the isolation of newly synthesized link protein and core protein precursor, as well as completed forms of the proteoglycan by taking advantage of their ability to be incorporated into aggregates recoverable from the bottom of CsCl density gradients. The second presents refinements of immunoprecipitation procedures, while the third describes the separation of two populations of newly synthesized aggregating proteoglycan by agarose gel electrophoresis. Bovine articular cartilage aggregating proteoglycans exhibit age-related differences. Electrophoresis in acrylamide-agarose composite gels has been used successfully to show that the matrix contains two populations of aggregating proteoglycans which are present in different ratios at different ages. Electrophoresis in 1% agarose gels has recently been found to offer a number of advantages over the acrylamideagarose composite gel procedure.