The role of ADAM-TS4 (aggrecanase-1) and ADAM-TS5 (aggrecanase-2) in a model of cartilage degradation
The role of ADAM-TS4 (aggrecanase-1) and ADAM-TS5 (aggrecanase-2) in a model of cartilage degradation
复制标题
DOI：
10.1053/joca.2001.0427
复制
发表时间：
2001-08-01
影响因子：
7
通讯作者：
Arner, E
中科院分区：
文献类型：
--
作者：
Tortorella, MD;Malfait, AM;Arner, E
简介：残基Glu（373）-Ala（374）之间聚集蛋白聚糖的切割被认为是关节炎疾病中聚集蛋白聚糖破坏的关键事件，其归因于酶活性，即聚集蛋白聚糖酶。已经鉴定了两种软骨聚集蛋白聚糖酶，聚集蛋白聚糖酶-1（ADAM-TS 4）和聚集蛋白聚糖酶-2（ADAM-TS 5）和两种酶已显示非常有效地在Glu（373）-Ala（374）位点切割可溶性聚集蛋白聚糖。确定ADAM-TS 4和/或ADAM-TS 5是否是负责白细胞介素-1（IL-1）和肿瘤坏死因子（TNF）后聚集蛋白聚糖催化的聚集蛋白聚糖酶。结果：（1）IL-1和TNF刺激下，聚集蛋白聚糖的释放与聚集蛋白聚糖在ADAM-TS 4和ADAM-TS 5敏感位点Glu（1480）Gly（1481）、Glu（1667）-Gly（1668）和Glu（1871）-Leu（1872）的C端断裂有关。(2)IL-1刺激软骨外植体后的切割顺序与用重组人ADAM-TS 4和ADAM-TS 5消化可溶性聚集蛋白聚糖时相同。(3)一般的金属蛋白酶抑制剂，但不是由MMP-特异性抑制剂，组成和刺激的切割的聚集蛋白聚糖在ADAM-TS 4和ADAM-TS 5敏感的网站在软骨中被阻止，这种抑制与抑制聚集蛋白聚糖从软骨中释放。(4)PCR和Western印迹分析表明，两种ADAM-TS蛋白酶在软骨外植体中表达; ADAM-TS 5组成型表达，而ADAM-TS 4在IL-1和TNF处理后被诱导。(5)IL-1刺激后，牛关节软骨培养物中ADAM-TS 4和ADAM-TS 5的免疫耗竭导致培养基中聚集蛋白聚糖酶活性降低90%。(C)2001年国际骨关节炎研究学会。
Introduction: Cleavage of aggrecan between residues Glu(373)-Ala(374), which is believed to be a key event in aggrecan destruction in arthritic diseases, has been attributed to an enzymatic activity, aggrecanase. Two cartilage aggrecanases have been identified, aggrecanase-1 (ADAM-TS4) and aggrecanase-2 (ADAM-TS5) and both enzymes have been shown very efficiently to cleave soluble aggrecan at the Glu(373)-Ala(374) site.Objective: To determine whether ADAM-TS4 and/or ADAM-TS5 are the aggrecanases responsible for aggrecan catabolism following interleukin-1 (IL-1) and tumor necrosis factor (TNF) treatment of bovine articular cartilage.Results: (1) IL-1- and TNF-stimulated release of aggrecan was associated with cleavage of aggrecan within the C-terminus at the ADAM-TS4 and ADAM-TS5-sensitive sites, Glu(1480)Gly(1481), Glu(1667)-Gly(1668) and Glu(1871)-Leu(1872). (2) The order of cleavage following IL-1 stimulation of cartilage explants; was the same as when soluble aggrecan is digested with recombinant human ADAM-TS4 and ADAM-TS5. (3) Both constitutive and stimulated cleavage of aggrecan at the ADAM-TS4 and ADAM-TS5-sensitive sites in cartilage was blocked by a general metalloproteinase inhibitor but not by a MMP-specific inhibitor, and this inhibition correlated with inhibition of aggrecan release from cartilage. (4) PCR and Western blot analysis indicated that both ADAM-TS proteases are expressed in cartilage explants; ADAM-TS5 is constitutively expressed whereas ADAM-TS4 is induced following IL-1 and TNF treatment. (5) Immunodepletion of both ADAM-TS4 and ADAM-TS5 from bovine articular cartilage cultures following IL-1 stimulation resulted in a 90% reduction of aggrecanase activity in the culture medium. (C) 2001 OsteoArthritis Research Society International.