CHARACTERIZATION OF A >400,000-DALTON CARTILAGE PROTEIN

各种非胶原性、非蛋白多糖蛋白质已在 关节软骨和非关节软骨。其中大部分的功能 蛋白质是未知的。我们最近发现了一种高分子量 (大于400,000道尔顿)蛋白质，共有116,000道尔顿亚基 研究了正常的关节软骨，以及正常的牛气管 软骨、萎缩的犬膝关节软骨和人类髋关节骨性关节炎 软骨。这种蛋白质约占总蛋白质的3-7%。 正常人4M氯化胍提取物中的非胶原蛋白 关节软骨是主要的非胶原蛋白之一 在正常犬膝关节软骨24小时器官培养中合成。 使用特定抗血清的免疫荧光研究显示， 116,000-Dalton蛋白在域间和细胞周围被发现 正常成人和胎儿软骨的区域。只有少量的 交叉反应物质存在于半月板、滑膜和滑膜细胞中。 文化。这种蛋白质不存在于血清或滑液中，这是 提供证据表明它没有从这些来源吸附到软骨上。 本申请中提出的研究的目标是提纯 未成熟关节软骨中含有大于40万道尔顿的蛋白质， 成熟的和骨关节炎的狗；为了探索它与胶原的相互作用， 蛋白多糖和透明质酸；在各种不同的 并研究其生化组成和代谢。这 因此，该项目将提供有关似乎是 正常关节软骨的一种主要的非胶原蛋白。它还可能 扩大我们对软骨大分子组成的了解 衰老和骨关节炎。

Various non-collagenous, non-proteoglycan proteins have been described in articular and non-articular cartilage. The function of most of these proteins is unknown. We have recently identified a high molecular weight (greater than 400,000-dalton) protein, with 116,000-dalton subunits, in all normal articular cartilage studied, as well as in normal bovine tracheal cartilage, atrophic canine knee cartilage, and osteoarthritic human hip cartilage. This protein constitutes approximately 3-7% of the total non-collagenous protein in 4 M guanidinium chloride extracts of normal articular cartilage and is one of the major non-collagenous proteins synthesized in 24-hours organ cultures of normal canine knee cartilage. Immunofluorescence studies using specific antiserum reveal that the 116,000-dalton protein is found in the interterritorial and pericellular regions of normal adult and fetal cartilage. Only a small amount of cross-reacting material is present in menisci, synovium, and synovial cell cultures. This protein is not present in serum or synovial fluid, which provides evidence that it is not adsorbed from these sources onto cartilage. The goals of the studies proposed in this application are to purify the greater than 400,000-dalton protein from articular cartilage of immature, mature, and osteoarthritic dogs; to explore its interactions with collagen, proteoglycans, and hyaluronic acid; to quantitate it in the various cartilages; and to study its biochemical composition and metabolism. This project will thus contribute new information concerning what appears to be a major non-collagenous protein of normal articular cartilage. It may also enlarge our understanding of the macromolecular composition of cartilage in aging and ostearthritis.