Biosynthesis of cartilage proteoglycan and link protein by articular chondrocytes from immature and mature rabbits.
Chondrocytes from immature and mature rabbits have been compared in biosynthetic studies with [3H] leucine and [35S]sulfate as precursors. The time course of incorporation of [3H]leucine into general protein, proteoglycan monomer core protein, and link protein and of [35S]sulfate into proteoglycan monomer has been examined. Proteoglycan monomer was isolated from the high buoyant density (p greater than 1.60) fractions of dissociative CsCl gradients and link protein by immunoprecipitation with antibody 8A4 followed by gel electrophoresis. Results based on the period of linear isotope incorporation showed that mature cells synthesize protein at about 40% of the rate of immature cells and both proteoglycan and link protein at about 20% of the rate of immature cells. The labeling rates obtained suggest that immature cells synthesize an approximate 1:1 molar ratio of link protein to proteoglycan monomer, and for mature cells this ratio is about 0.8:1. While cell layer retention of newly synthesized proteoglycan was markedly lower in mature relative to immature cell cultures, link protein retention was high in both immature and mature cultures; this finding provides an explanation for our previous observation (Plaas, A. H. K., and Sandy, J. D. (1984) Biochem, J. 220, 337-340) that link-free monomer accumulates in the medium of mature but not immature cultures. The link protein synthesized by both ages of cells and isolated from cell layer or medium was a single major species of apparent molecular mass 48-51 kDa. The results suggest that mature chondrocytes are less efficient than immature chondrocytes in the coordinated assembly of link-stabilized proteoglycan aggregates in this culture system.
Plaas
A H
AH
Department of Orthopaedics, Brown University, Rhode Island Hospital, Providence 02906.
Sandy
J D
JD
Kimura
J H
JH
D000818:Animals; D002358:Cartilage, Articular / Q000378:metabolism*; D002478:Cells, Cultured; D016326:Extracellular Matrix Proteins; D007930:Leucine / Q000378:metabolism; D008970:Molecular Weight; D014176:Protein Biosynthesis; D011509:Proteoglycans / Q000096:biosynthesis*; D011817:Rabbits; D013431:Sulfates / Q000378:metabolism; D013997:Time Factors; D000099249:Proteoglycan Link Protein
D016326:Extracellular Matrix Proteins; D007930:Leucine; D011509:Proteoglycans; D013431:Sulfates; D000099249:Proteoglycan Link Protein