Demonstration of epidermal growth factor-induced receptor dimerization in living cells using a chemical covalent cross-linking agent.
Demonstration of epidermal growth factor-induced receptor dimerization in living cells using a chemical covalent cross-linking agent.
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DOI：
10.1016/s0021-9258(18)69070-2
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发表时间：
1988-03
期刊：
影响因子：
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通讯作者：
C. Cochet;O. Kashles;E. Chambaz;I. Borrello;C. King;J. Schlessinger
中科院分区：
文献类型：
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作者：
C. Cochet;O. Kashles;E. Chambaz;I. Borrello;C. King;J. Schlessinger
我们用可溶性共价交联剂1-乙基-3-[3-（二甲氨基）丙基]碳二亚胺（EDAC）研究了表皮生长因子（EGF）刺激纯化的EGF受体、膜制备物和完整的A431细胞中EGF受体二聚化的能力。将EGF加入到来自A431细胞的膜上或加入到通过免疫亲和层析从A431细胞纯化的EGF受体上，导致出现Mr 340，000的交联产物，该交联产物用EGF受体特异性抗体鉴定为EGF受体二聚体。三个独立的方法，包括生物合成标记，表面碘化，和免疫印迹实验被用来跟踪EGF受体二聚体在活的A431细胞。这些方法提供了一致的结果，表明EGF诱导活细胞中EGF受体的快速二聚化，表明该过程可能在EGF介导的跨膜信号传导中起作用。
We have used the soluble covalent cross-linking agent 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDAC) to examine the capacity of epidermal growth factor (EGF) to stimulate the dimerization of purified EGF receptor, of EGF receptor in membrane preparations and in intact A431 cells. The addition of EGF either to membranes from A431 cells or to EGF receptor which was purified from A431 cells by immunoaffinity chromatography caused the appearance of a cross-linked product of Mr 340,000 which was identified using EGF receptor-specific antibodies as an EGF receptor dimer. Three independent approaches including biosynthetic labeling, surface iodination, and immunoblotting experiments were utilized to follow EGF receptor dimerization in living A431 cells. These approaches provided consistent results indicating that EGF induced rapid dimerization of EGF receptor in living cells, suggesting that this process may play a role in transmembrane signalling mediated by EGF.