FHL2 interacts with both ADAM-17 and the cytoskeleton and regulates ADAM-17 localization and activity
FHL2 interacts with both ADAM-17 and the cytoskeleton and regulates ADAM-17 localization and activity
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DOI：
10.1002/jcp.20671
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发表时间：
2006-08-01
影响因子：
5.6
通讯作者：
Peiretti, Franck
中科院分区：
文献类型：
--
作者：
Canault, Matthias;Tellier, Edwige;Peiretti, Franck
ADAM - 17是一种金属蛋白酶解聚素，负责几种跨膜蛋白的胞外结构域脱落。利用酵母双杂交系统，我们发现ADAM - 17与四个半LIM结构域2蛋白（FHL2）相互作用，FHL2是一种参与多种蛋白质 - 蛋白质相互作用的LIM结构域蛋白。我们证明这种相互作用涉及ADAM - 17从第721位到第739位的氨基酸序列。在心肌成肌细胞H9C2中，ADAM - 17和FHL2与基于肌动蛋白的细胞骨架共定位，并且我们发现FHL2与ADAM - 17和基于肌动蛋白的细胞骨架都结合。我们发现主要是ADAM - 17的成熟形式与细胞骨架结合，尽管弗林蛋白酶对ADAM - 17的成熟作用对于其与细胞骨架的结合不是必需的。有趣的是，与FHL2缺陷型巨噬细胞相比，在野生型小鼠巨噬细胞表面检测到的ADAM - 17较少。然而，在佛波醇酯（PMA）刺激下，野生型细胞释放ADAM - 17底物的能力更强。总之，这些结果证明了ADAM - 17和FHL2之间存在物理和功能上的相互作用，这意味着FHL2在ADAM - 17的调节中起作用。
ADAM-17 is a metalloprotease-disintegrin responsible for the ectodomain shedding of several transmembrane proteins. Using the yeast two-hybrid system, we showed that ADAM-17 interacts with the Four and Half LIM domain 2 protein (FHL2), a LIM domain protein that is involved in multiple protein-protein interaction. We demonstrated that this interaction involved the amino-acid sequence of ADAM-17 from position 721 to 739. In the cardiomyoblast cells H9C2, ADAM-17 and FHL2 colocalize with the actin-based cytoskeleton and we showed that FHL2 binds both ADAM-17 and the actin-based cytoskeleton. We found that mainly the mature form of ADAM-17 associates with the cytoskeleton, although the maturation of ADAM-17 by furin is not necessary for its binding to the cytoskeleton. Interestingly, less ADAM-17 was detected at the surface of wild-type mouse macrophages compared to FHL2 deficient macrophages. However, wild-type cells have a higher ability to release ADAM-17 substrates under PMA stimulation. Altogether, these results demonstrate a physical and functional interaction between ADAM-17 and FHL2 that implies that FHL2 has a role in the regulation of ADAM-17.