The EGF receptor is an actin-binding protein.
The EGF receptor is an actin-binding protein.
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DOI：
10.1083/jcb.119.2.349
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发表时间：
1992-10
期刊：
影响因子：
--
通讯作者：
Boonstra J
中科院分区：
文献类型：
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作者：
den Hartigh JC;van Bergen en Henegouwen PM;Verkleij AJ;Boonstra J
在最近的一些研究中，已经表明，在体内部分的表皮生长因子受体（EGFR）的人口是与肌动蛋白丝系统。在本文中，我们证明了纯化的EGFR可以与纯化的丝状肌动蛋白（F-肌动蛋白）共沉淀，表明EGFR和肌动蛋白之间的直接关联。一个截短的EGFR，以前被证明是不相关的细胞骨架，被用作对照，这种受体没有共沉淀与肌动蛋白丝。通过测量多克隆抗体或合成肽对EGFR与F-肌动蛋白共沉降的竞争来确定EGFR的肌动蛋白结合结构域。合成的肽与EGFR的氨基酸残基984-996（HL-33）同源，其显示与阿米巴profilin的肌动蛋白结合结构域的高度同源性。发现针对HL-33的多克隆抗体可防止EGFR与F-肌动蛋白的共沉淀。该肽HL-33显示出直接与肌动蛋白结合，而与残基1001-1013（HL-34）同源的合成肽相反。在共沉淀过程中，HL-33竞争EGFR的肌动蛋白结合，而HL-34不竞争，表明EGFR含有一个肌动蛋白结合位点。这些结果表明EGFR是肌动蛋白结合蛋白，其通过含有氨基酸残基984-996的结构域与肌动蛋白结合。
In a number of recent studies it has been shown that in vivo part of the EGF receptor (EGFR) population is associated to the actin filament system. In this paper we demonstrate that the purified EGFR can be cosedimented with purified filamentous actin (F-actin) indicating a direct association between EGFR and actin. A truncated EGFR, previously shown not to be associated to the cytoskeleton, was used as a control and this receptor did not cosediment with actin filaments. Determination of the actin-binding domain of the EGFR was done by measuring competition of either a polyclonal antibody or synthetic peptides on EGFR cosedimentation with F-actin. A synthetic peptide was made homologous to amino acid residues 984-996 (HL-33) of the EGFR which shows high homology with the actin-binding domain of Acanthamoeba profilin. A polyclonal antibody raised against HL-33 was found to prevent cosedimentation of EGFR with F-actin. This peptide HL-33 was shown to bind directly to actin in contrast with a synthetic peptide homologous to residues 1001-1013 (HL-34). During cosedimentation, HL-33 competed for actin binding of the EGFR and HL-34 did not, indicating that the EGFR contains one actin-binding site. These results demonstrate that the EGFR is an actin-binding protein which binds to actin via a domain containing amino acids residues 984-996.