表皮生长因子受体的两个结构域参与细胞骨架相互作用。

Two domains of the epidermal growth factor receptor are involved in cytoskeletal interactions.

作者信息

Song Wei, Wu Jing, Ge Gaoxiang, Lin Qishui

机构信息

Key Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.

出版信息

Biochem Biophys Res Commun. 2008 Jun 13;370(4):589-93. doi: 10.1016/j.bbrc.2008.03.138. Epub 2008 Apr 7.

DOI:10.1016/j.bbrc.2008.03.138

PMID:18396150

Abstract

Epidermal growth factor receptor can interact directly with F-actin through an actin-binding domain. In the present study, a mutant EGFR, lacking a previously identified actin-binding domain (ABD 1), was still able to bind elements of the cytoskeleton. A second EGFR actin-binding domain (ABD 2) was identified in the region of the receptor that includes Tyr-1148 by a yeast two-hybrid assay. GST fusion proteins comprising ABD 1 or ABD 2 bound actin in vitro and competed for actin-binding with the full-length EGFR. EGFR binding to actin was also studied in intact cells using fluorescence resonance energy transfer (FRET). The localization of the EGFR/actin-binding complex changed after EGF stimulation. Fusion proteins containing mutations in ABD1 or ABD2 did not display a FRET signal. The results lead to the conclusion that the interaction between ABD1 and ABD2 and actin during EGF-induced signal transduction, and thus between EGFR and actin, are important in cell activation.

摘要

表皮生长因子受体可通过一个肌动蛋白结合结构域直接与F-肌动蛋白相互作用。在本研究中，一个缺乏先前鉴定出的肌动蛋白结合结构域（ABD 1）的突变型表皮生长因子受体（EGFR）仍能够结合细胞骨架成分。通过酵母双杂交试验在受体包含酪氨酸-1148的区域鉴定出了第二个EGFR肌动蛋白结合结构域（ABD 2）。包含ABD 1或ABD 2的谷胱甘肽S-转移酶（GST）融合蛋白在体外结合肌动蛋白，并与全长EGFR竞争肌动蛋白结合。还利用荧光共振能量转移（FRET）在完整细胞中研究了EGFR与肌动蛋白的结合。表皮生长因子（EGF）刺激后，EGFR/肌动蛋白结合复合物的定位发生了变化。在ABD1或ABD2中含有突变的融合蛋白未显示出FRET信号。结果得出结论，在EGF诱导的信号转导过程中，ABD1和ABD2与肌动蛋白之间的相互作用，进而EGFR与肌动蛋白之间的相互作用，在细胞激活中很重要。