mGrb10 interacts with Nedd4
mGrb10 interacts with Nedd4
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DOI：
10.1074/jbc.274.34.24094
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发表时间：
1999-08-20
影响因子：
4.8
通讯作者：
Baserga, R
中科院分区：
文献类型：
--
作者：
Morrione, A;Plant, P;Baserga, R
我们利用酵母双杂交系统来鉴定与小鼠 Grb10 相互作用的蛋白质，Grb10 是一种已知与胰岛素和胰岛素样生长因子 I 受体相互作用的衔接蛋白。我们分离了一个小鼠 cDNA 克隆，其中包含小鼠 Nedd4 的 C2 结构域、泛素蛋白连接酶 (E3)，还包含一个甜菜结构域（与 EG-AP 羧基末端同源）和三个 WW 结构域。使用全长 Nedd4 证实了双杂交系统中与 Grb10 的相互作用，并通过删除 Grb10 的最后 148 个氨基酸（包含 SH2 结构域和新鉴定的 BPS 结构域的区域）而消除了相互作用。 Grb10 和 Nedd4 之间的相互作用也在小鼠胚胎成纤维细胞中重现，其中内源性 Nedd4 与内源性和过表达的 Grb10 发生组成型共免疫沉淀。这种相互作用不依赖于 Ca2+。 Grb10 与 Nedd4 的相互作用在体内并未被泛素化，这增加了这种相互作用可用于靶向其他蛋白质（如酪氨酸激酶受体）进行泛素化的可能性。
We have utilized the yeast two-hybrid system to identify proteins interacting with mouse Grb10, an adapter protein known to interact with both the insulin and the insulin-like growth factor-I receptors. We have isolated a mouse cDNA clone containing the C2 domain of mouse Nedd4, a ubiquitin protein ligase (E3) that also contains a beet (homologous to the EG-AP carboxyl-terminus) domain and three WW domains. The interaction with Grb10 in the two-hybrid system was confirmed using the full-length Nedd4, and it was abolished by deleting the last 148 amino acids of Grb10, a region that includes the SH2 domain and the newly identified BPS domain. The interaction between Grb10 and Nedd4 was also reproduced in vivo in mouse embryo fibroblasts, where endogenous Nedd4 co-immunoprecipitated constitutively with both the endogenous and an overexpressed Grb10. This interaction was Ca2+-independent. Grb10 interacting with Nedd4 was not ubiquitinated in vivo, raising the possibility that this interaction may be used to target other proteins, like tyrosine kinase receptors, for ubiquitination.