THE STRUCTURE OF AGGRECAN FRAGMENTS IN HUMAN SYNOVIAL-FLUID - EVIDENCE FOR THE INVOLVEMENT IN OSTEOARTHRITIS OF A NOVEL PROTEINASE WHICH CLEAVES THE GLU-373-ALA-374 BOND OF THE INTERGLOBULAR DOMAIN
THE STRUCTURE OF AGGRECAN FRAGMENTS IN HUMAN SYNOVIAL-FLUID - EVIDENCE FOR THE INVOLVEMENT IN OSTEOARTHRITIS OF A NOVEL PROTEINASE WHICH CLEAVES THE GLU-373-ALA-374 BOND OF THE INTERGLOBULAR DOMAIN
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DOI：
10.1172/jci115742
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发表时间：
1992-05-01
影响因子：
15.9
通讯作者：
LOHMANDER, LS
中科院分区：
文献类型：
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作者：
SANDY, JD;FLANNERY, CR;LOHMANDER, LS
从近期膝关节损伤患者和早期或晚期骨关节炎患者中收集滑液。硫酸软骨素取代的聚集蛋白聚糖片段存在于这些液体中，并在正常牛滑液中，通过氯化铯梯度离心纯化，酶促去糖基化和分级凝胶过滤Superose-12。每个样品含有两个主要的聚集蛋白聚糖核心蛋白群体，表观分子量约为90 kD和150 kD。对于所有样品，两个群体的NH 2-末端分析得到了一个单一的主要序列开始ARGSV。该NH 2末端由人聚集蛋白聚糖核心蛋白在G1和G2结构域之间的球间结构域内的Glu 373-Ala 374键处的切割产生。在牛软骨外植体培养物中，在对照和白细胞介素-1刺激的聚集蛋白聚糖催化过程中也发生在该位点的切割（桑迪，J.，尼姆河Boynton和C.弗兰纳里1991.这些结果表明，骨关节炎人滑液和正常牛滑液中存在的主要聚集蛋白聚糖片段都很大，由球间结构域、G2结构域、硫酸角质素结构域和可变长度的硫酸软骨素结构域的短NH 2末端延伸组成。我们得出结论，聚集蛋白聚糖片段从关节软骨释放到滑液中见于人骨关节炎的所有阶段（Lohmander，L. S. 1991.骨科学报Scand.62：623-632）是由正常软骨蛋白酶的作用促进的，所述蛋白酶切割球间结构域的Glu 373-Ala 374键。
Synovial fluid was collected from patients with recent knee injury and from patients with early or late stage osteoarthritis. Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. For all samples, NH2-terminal analysis of both populations gave a single major sequence beginning ARGSV. This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1991. J. Biol. Chem. 266:8683-8685).These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. Acta Orthop. Scand. 62:623-632) is promoted by the action of a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain.