Characterization of METH-1/ADAMTS1 processing reveals two distinct active forms
Characterization of METH-1/ADAMTS1 processing reveals two distinct active forms
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DOI：
10.1074/jbc.m002599200
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发表时间：
2000-10-27
影响因子：
4.8
通讯作者：
Iruela-Arispe, ML
中科院分区：
文献类型：
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作者：
Rodríguez-Manzaneque, JC;Milchanowski, AB;Iruela-Arispe, ML
METH-1/ADAMTS 1是一个新发现的含有金属蛋白酶、去整合素和血小板反应蛋白样基序的基因家族的成员。我们最近发现METH-1蛋白是一种有效的血管生成抑制剂。在这里，我们证明分泌的人pro-METH-1在两个连续的步骤中被处理以释放p87和p65活性形式。p87形式缺乏N-末端前结构域，p65由C-末端的额外加工事件产生。p87的产生被弗林蛋白酶的特异性抑制剂阻断，并且pro-METH-1与纯化的弗林蛋白酶一起孵育释放p87片段而不是p65。p65的产生需要p87的预形成并且被基质金属蛋白酶的抑制剂抑制。我们证明，基质金属蛋白酶2，8和15能够释放p65时，p87被用作底物。这第二个处理步骤从p87-METH-1的羧基末端去除两个血小板反应蛋白重复序列，并改变蛋白质对肝素和内皮培养物的亲和力。此外，这种缺失与抑制内皮细胞增殖后活性降低有关。我们假设METH-1加工与调节该蛋白质显示的抗血管生成特性相关。
METH-1/ADAMTS1 is a member of a newly described family of genes that contain metalloprotease, disintegrin, and thrombospondin-like motifs. We have recently shown that METH-1 protein is a potent inhibitor of angiogenesis, Here, we demonstrate that secreted human pro-METH-1 is processed in two consecutive steps to release both p87 and p65 active forms. The p87 form lacks the N-terminal prodomain and p65 results from an additional processing event in the C-terminal end. Generation of p87 was blocked with specific inhibitors of furin, and incubation of pro-METH-1 with purified furin released the p87 fragment but not p65, Generation of p65 required preformation of p87 and was suppressed by inhibitors of matrix metalloproteases. We demonstrate that matrix metalloproteases 2, 8, and 15 were able to release p65 when p87 was used as substrate. This second processing step removes two thrombospondin repeats from the carboxyl-terminal end of p87-METH-1 and alters the affinity of the protein to heparin and endothelial cultures. Furthermore, this deletion was associated with a reduced activity upon suppression of endothelial cell proliferation, We hypothesize that METH-1 processing is relevant for the modulation of the anti-angiogenic properties displayed by the protein.