The adaptor protein fish associates with members of the ADAMs family and localizes to podosomes of Src-transformed cells
The adaptor protein fish associates with members of the ADAMs family and localizes to podosomes of Src-transformed cells
复制标题
DOI：
10.1074/jbc.m300267200
复制
发表时间：
2003-05-09
影响因子：
4.8
通讯作者：
Courtneidge, SA
中科院分区：
文献类型：
--
作者：
Abram, CL;Seals, DF;Courtneidge, SA
Fish是一种支架蛋白和Src底物。它含有一个氨基末端Phox同源（PX）结构域和五个Src同源3（SH 3）结构域，以及用于结合含SH 2和SH 3结构域的蛋白质的多个基序。我们已经确定，鱼的PX结构域结合3-磷酸化磷脂酰肌醇（包括磷脂酰肌醇3-磷酸和磷脂酰肌醇3，4-二磷酸）。与此一致，绿色荧光蛋白和鱼PX结构域的融合蛋白定位于与正常成纤维细胞中的内体类似的点状结构。然而，全长鱼蛋白主要是细胞质，这表明其PX结构域可能无法在未刺激的细胞中进行分子间相互作用。在Src转化的细胞中，我们观察到一个戏剧性的重新定位的一些鱼分子肌动蛋白丰富的结构称为podosomes的PX结构域是必要的和足够的影响这种易位。我们使用噬菌体展示屏幕与第五SH 3域的鱼和分离的ADAM 19作为结合伴侣。随后在哺乳动物细胞中的分析表明，鱼与亚当斯家族的几个成员相互作用，包括亚当斯12，15和19。在Src转化的细胞中，ADAM 12与Fish共定位于podosomes中。由于亚当斯家族的成员与生长因子加工以及细胞粘附和运动有关，因此Fish可能充当允许Src冲击这些过程的适配器分子。
Fish is a scaffolding protein and Src substrate. It contains an amino-terminal Phox homology (PX) domain and five Src homology 3 (SH3) domains, as well as multiple motifs for binding both SH2 and SH3 domain-containing proteins. We have determined that the PX domain of Fish binds 3-phosphorylated phosphatidylinositols (including phosphatidylinositol 3-phosphate and phosphatidylinositol 3,4-bisphosphate). Consistent with this, a fusion protein of green fluorescent protein and the Fish PX domain localized to punctate structures similar to endosomes in normal fibroblasts. However, the full-length Fish protein was largely cytoplasmic, suggesting that its PX domain may not be able to make intermolecular interactions in unstimulated cells. In Src-transformed cells, we observed a dramatic re-localization of some Fish molecules to actin-rich structures called podosomes; the PX domain was both necessary and sufficient to effect this translocation. We used a phage display screen with the fifth SH3 domain of Fish and isolated ADAM19 as a binding partner. Subsequent analyses in mammalian cells demonstrated that Fish interacts with several members of the ADAMs family, including ADAMs 12, 15, and 19. In Src-transformed cells, ADAM12 co-localized with Fish in podosomes. Because members of the ADAMs family have been implicated in growth factor processing, as well as cell adhesion and motility, Fish could be acting as an adaptor molecule that allows Src to impinge on these processes.