Characterization of catabolin, the major product of pig synovial tissue that induces resorption of cartilage proteoglycan in vitro.
Characterization of catabolin, the major product of pig synovial tissue that induces resorption of cartilage proteoglycan in vitro.
复制标题
分解代谢蛋白的表征，分解代谢蛋白是猪滑膜组织的主要产物，可在体外诱导软骨蛋白多糖的吸收。
作者：
J. Saklatvala
1.器官培养中的猪滑膜产生诱导活软骨在体外吸收其蛋白聚糖的物质。2.该材料的生物测定是测量培养8天的牛鼻中隔软骨外植体释放的糖胺聚糖。通过加入琥珀酸皮质醇（0.1 μ g/ml），测定的性能大大提高。这减少了未受刺激的软骨中糖胺聚糖的释放，而不抑制其对滑膜分解代谢因子的反应。3.通过使用这种改进的测定，显示滑膜培养基中90%的活性物质被透析膜保留。4.通过（NH（4））（2）SO（4）沉淀、离子交换层析、凝胶过滤和制备等电聚焦，从滑膜培养液中部分纯化了活性蛋白。5.这种蛋白质，称为代谢产物，具有分子量。17000和pI4.6。6.将在透析管中浓缩的滑膜培养基进行凝胶色谱，发现含有一种主要活性组分，其在与部分纯化的分解代谢产物相同的位置洗脱。7.滑液培养基不能通过加热（70 ℃，10分钟）灭活，部分纯化的分解代谢产物的稀释制剂也不能灭活，但浓缩的粗制剂是热不稳定的。8.这些结果表明，分解代谢素是体外培养的滑膜组织诱导活软骨蛋白多糖吸收的主要物质。9.其他培养的软结缔组织产生分解代谢素样活性。以巩膜为例，琥珀酸皮质醇（0.1 μ g/ml）可抑制其产生。10.提示分解代谢素可能是软结缔组织在培养过程中的一般产物，其生理功能可能是诱导结缔组织基质在损伤后的吸收。
1. Pig synovium in organ culture produces material which induces living cartilage to resorb its proteoglycan in vitro. 2. The bioassay for this material was to measure glycosaminoglycan released from explants of bovine nasal-septal cartilage cultured for 8 days. The performance of the assay was greatly improved by adding cortisol succinate (0.1mug/ml). This decreased the release of glycosaminoglycan from unstimulated cartilage without inhibiting its response to catabolic factors from the synovium. 3. By using this improved assay it was shown that 90% of the active materials in synovial culture medium were retained by dialysis membrane. 4. An active protein was partially purified from synovial culture medium by (NH(4))(2)SO(4) precipitation, ion-exchange chromatography, gel filtration and preparative isoelectric focusing. 5. This protein, called catabolin, had mol.wt. 17000 and pI4.6. 6. Synovial culture medium concentrated in dialysis tubing was subjected to gel chromatography and found to contain one major active component, which was eluted at the same position as the partially purified catabolin. 7. The synovial culture medium was not inactivated by heating (70 degrees C for 10min), nor were diluted preparations of partially purified catabolin, but concentrated crude preparations were thermolabile. 8. These results suggest that catabolin is the major substance produced by the synovial tissue in culture which induces resorption of proteoglycan of living cartilage in vitro. 9. Other cultured soft connective tissues produced catabolin-like activity. The example of sclera is shown, and production was inhibited by cortisol succinate (0.1mug/ml). 10. It is suggested that catabolin may be a general product of soft connective tissues in culture, and its physiological function may be to induce resorption of connective-tissue matrix after injury.