THE EFFECT OF MATURATION AND AGING ON THE STRUCTURE AND CONTENT OF LINK PROTEINS IN RABBIT ARTICULAR-CARTILAGE

We have examined extracts of articular cartilage from rabbits aged 3–100 weeks for evidence of age‐related changes in the structure and content of link protein (LP) in this tissue, with the following findings: (a) Two major molecular weight forms of LP were seen on SDS–PAGE (41 and 48 kDa) and the proportion of these changed markedly with age. The 48 kDa species was predominant in young animals (representing about 78% of the total LP at 5 weeks) whereas the 41 kDa species increased in amount with age (representing 35% of the total LP at 100 weeks). A minor form of about 43 kDa, representing less than 20% of the total, was present only during the growth phase. A small amount of fragmented link protein (less than 5% of the total) of about 25–30 kDa was present in samples from mature and aged rabbits only. (b) The quantitation of LP in guanidinium: HCl extracts of cartilage, by radioimmunoassay with monoclonal antibody 8‐A‐4, was markedly influenced by the conditions of preparation and pretreatment of samples. Assays of dialyzed guanidine extracts following treatment at 80°C for 15 min in 0.025% (w/v) SDS indicated that immature and mature cartilage contains about 50 and 180 μg of LP/g of tissue, respectively. On the other hand, assays following treatment at 100°C for 20 min in 0.1% (w/v) SDS suggested that rabbit cartilage contains about 300 μg of LP/g of tissue at all ages; finally, assay of CsCl purified proteoglycan samples under these conditions indicated a content of about 500 μg of LP/g at all ages. (c) Calculations based on the analysis of proteoglycan preparations for aggregating monomer and link protein suggest that a LP:aggregating monomer molar ratio of about 0.9 is maintained in the articular cartilage throughout maturation and aging in the rabbit. Copyright © 1990 Orthopaedic Research Society