Human Intersectin 2 (ITSN2) binds to Eps8 protein and enhances its degradation
Human Intersectin 2 (ITSN2) binds to Eps8 protein and enhances its degradation
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Human Intersectin 2 (ITSN2) 与 Eps8 蛋白结合并增强其降解
DOI：
10.5483/bmbrep.2012.45.3.183
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发表时间：
2012-03-31
期刊：
影响因子：
3.8
通讯作者：
Zhang, Jian
中科院分区：
文献类型：
--
作者：
Ding, Xiaofeng;Yang, Zijian;Zhang, Jian
通过Rac参与肌动蛋白重塑，通过Rab5参与受体内吞。在这里，我们使用酵母双杂交系统，以Eps8为诱饵来筛选人脑cDNA文库。ITSN2被鉴定为Eps8的新结合因子。通过体内免疫共沉淀和共定位实验以及体外GST下拉实验证实了ITSN2和Eps8之间的相互作用。此外，我们还将相互作用结构域定位到EPS8的260-306位氨基酸与ITSN2的盘绕线圈结构域之间。此外，蛋白质稳定性分析和免疫荧光分析表明，ITSN2过表达诱导了Eps8蛋白的降解，溶酶体抑制剂NH4Cl处理后这种降解作用明显减轻。综上所述，我们的结果表明，ITSN2与Eps8相互作用，并刺激Eps8蛋白的降解。[BMB报告2012；45(3)：183-188]
Participates in actin remodeling through Rac and receptor endocytosis via Rab5. Here, we used yeast two-hybrid system with Eps8 as bait to screen a human brain cDNA library. ITSN2 was identified as the novel binding factor of Eps8. The interaction between ITSN2 and Eps8 was demonstrated by the in vivo co-immunoprecipitation and colocalization assays and the in vitro GST pull-down assays. Furthermore, we mapped the interaction domains to the region between amino acids 260-306 of Eps8 and the coiled-coil domain of ITSN2. In addition, protein stability assays and immunofluorescence analysis showed ITSN2 overexpression induced the degradation of Eps8 proteins, which was markedly alleviated with the lysosome inhibitor NH4Cl treatment. Taken together, our results suggested ITSN2 interacts with Eps8 and stimulates the degradation of Eps8 proteins. [BMB reports 2012; 45(3): 183-188]