Integrin α5β1 and ADAM-17 interact in vitro and co-localize in migrating HeLa cells
Integrin α5β1 and ADAM-17 interact in vitro and co-localize in migrating HeLa cells
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DOI：
10.1074/jbc.m400180200
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发表时间：
2004-05-21
影响因子：
4.8
通讯作者：
Humphries, MJ
中科院分区：
文献类型：
--
作者：
Bax, DV;Messent, AJ;Humphries, MJ
肿瘤坏死因子（TNF）α转化酶（TACE/ADAM-17）在细胞表面分子的蛋白水解加工中具有多种作用，并且由于其加工TNF α的能力，是抗炎疗法的经验证的治疗靶标。与许多其他ADAM蛋白不同，其通过其去整合素结构域与整联蛋白受体相互作用，目前没有证据表明ADAM-17-整联蛋白缔合。通过分析一系列细胞系与ADAM-17胞外结构域重组片段的粘附，我们现在证明了ADAM-17和α（5）β（1）整合素之间的反式相互作用。由于ADAM-17介导的粘附对RGD肽和EDTA敏感，并且ADAM-17内的整合素结合位点缩小到去整合素/富含半胱氨酸的区域，因此这两种分子似乎具有由α（5）β（1）配体结合口袋介导的配体-受体关系。有趣的是，ADAM-17和alpha（5）beta（1）被发现共同定位于HeLa细胞的膜皱褶和粘着斑中。当融合的HeLa细胞单层被损伤时，ADAM-17和α（5）β（1）重新分布到前缘并共定位，这表明顺式取向。我们推测ADAM-17与alpha（5）beta（1）的相互作用可能靶向或调节其金属蛋白水解活性。
Tumor necrosis factor (TNF) alpha-converting enzyme (TACE/ADAM-17) has diverse roles in the proteolytic processing of cell surface molecules and, due to its ability to process TNFalpha, is a validated therapeutic target for anti-inflammatory therapies. Unlike a number of other ADAM proteins, which interact with integrin receptors via their disintegrin domains, there is currently no evidence for an ADAM-17-integrin association. By analyzing the adhesion of a series of cell lines with recombinant fragments of the extracellular domain of ADAM-17, we now demonstrate a functional interaction between ADAM-17 and alpha(5)beta(1) integrin in a trans orientation. Because ADAM-17-mediated adhesion was sensitive to RGD peptides and EDTA, and the integrin-binding site within ADAM-17 was narrowed down to the disintegrin/cysteine-rich region, the two molecules appear to have a ligand-receptor relationship mediated by the alpha(5)beta(1) ligand binding pocket. Intriguingly, ADAM-17 and alpha(5)beta(1) were found to co-localize in both membrane ruffles and focal adhesions in HeLa cells. When confluent HeLa cell monolayers were wounded, ADAM-17 and alpha(5)beta(1) redistributed to the leading edge and co-localized, which is suggestive of a cis orientation. We postulate that the interaction of ADAM-17 with alpha(5)beta(1) may target or modulate its metalloproteolytic activity.