Cloning and characterization of ADAMTS11, an aggrecanase from the ADAMTS family
Cloning and characterization of ADAMTS11, an aggrecanase from the ADAMTS family
复制标题
DOI：
10.1074/jbc.274.33.23443
复制
发表时间：
1999-08-13
影响因子：
4.8
通讯作者：
Burn, TC
中科院分区：
文献类型：
--
作者：
Abbaszade, I;Liu, RQ;Burn, TC
Aggrecan负责软骨的机械性能。在关节炎中观察到的最早的变化之一是由于蛋白水解裂解的增加而导致软骨脂肪的消耗。在ASN（341）-phe（342）和Glu（373）-Ala（374）的ASN（341）-Phe（342）和GLU（374）的该区域中，已经确定了两个主要的裂解部位，而已显示几种基质金属蛋白酶在ASN（341）-Phe（342）-phe（342）裂解，一种尚未识别的蛋白质称为“ aggrecanase”，负责乳沟Glu（373）-Ala（374），并假设在软骨损伤中起关键作用。我们已经鉴定并克隆了一种新型的拆分蛋白金属蛋白酶，具有血小板蛋白基序，具有ADAMTS11（Adamts11（Aggrecanase-2）），与ADAMTS4（Adamts4（Aggrecanase-1）具有广泛的同源性）已显示在整合素结合中起作用，细胞 - 细胞相互作用和细胞外基质结合。我们已经在昆虫细胞中表达了重组人ADAMTS11，并表明它在GLU（373）-ALA（374）位点裂解了Aggrecan，其裂解模式和抑制剂曲线与与天然Aggrcanase观察到的相比，其结构和结构和结构和结构和结构和结构和结构和抑制还描述了ADAMTS11，ADAMTS4和ADAMTS1的表达模式。我们的发现将促进软骨降解机制的研究，并为寻找有效的关节炎软骨耗竭抑制剂提供了靶标。
Aggrecan is responsible for the mechanical properties of cartilage. One of the earliest changes observed in arthritis is the depletion of cartilage aggrecan due to increased proteolytic cleavage within the interglobular domain. Two major sites of cleavage have been identified in this region at Asn(341)-Phe(342) and Glu(373)-Ala(374) While several matrix metalloproteinases have been shown to cleave at Asn(341)-Phe(342), an as yet unidentified protein termed "aggrecanase" is responsible for cleavage at Glu(373)-Ala(374) and is hypothesized to play a pivotal role in cartilage damage. We have identified and cloned a novel disintegrin metalloproteinase with thrombospondin motifs that possesses aggrecanase activity, ADAMTS11 (aggrecanase-2), which has extensive homology to ADAMTS4 (aggrecanase-1) and the inflammation-associated gene ADAMTS1, ADAMTS11 possesses a number of conserved domains that have been shown to play a role in integrin binding, cell-cell interactions, and extracellular matrix binding. We have expressed recombinant human ADAMTS11 in insect cells and shown that it cleaves aggrecan at the Glu(373)-Ala(374) site, with the cleavage pattern and inhibitor profile being indistinguishable from that observed with native aggrecanase, A comparison of the structure and expression patterns of ADAMTS11, ADAMTS4, and ADAMTS1 is also described. Our findings will facilitate the study of the mechanisms of cartilage degradation and provide targets to search for effective inhibitors of cartilage depletion in arthritic disease.