EPIDERMAL GROWTH-FACTOR STIMULATES PHOSPHORYLATION IN MEMBRANE PREPARATIONS INVITRO
EPIDERMAL GROWTH-FACTOR STIMULATES PHOSPHORYLATION IN MEMBRANE PREPARATIONS INVITRO
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DOI：
10.1038/276409a0
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发表时间：
1978-01-01
期刊：
影响因子：
64.8
通讯作者：
COHEN, S
中科院分区：
文献类型：
--
作者：
CARPENTER, G;KING, L;COHEN, S
表皮生长因子（EGF）与完整细胞中的质膜受体形成复合物，启动一系列生物化学事件，导致体内和体外细胞生长增加1。EGF与膜受体的相互作用已在粗膜纯化中得到证实2，但未检测到激素结合引起的膜的生化改变。为了阐明调节细胞增殖的分子机制，需要在无细胞系统中研究由有丝分裂原如EGF引发的特定生化反应。由于人表皮样癌细胞系A-431具有非常高浓度的EGF受体3，4（每个细胞2-3 × 106个受体），我们使用来自这些细胞的粗制膜制剂来寻找膜结构和/或功能的EGF依赖性改变。本文报道：（1）用能特异结合~（125）I标记的EGF的A-431细胞制备细胞膜;（2）EGF与这些细胞膜的结合，在[γ-32 P]ATP存在下，能显著刺激内源性蛋白质的磷酸化。
EPIDERMAL GROWTH FACTOR (EGF) forms a complex with plasma membrane receptors in intact cells that initiates a series of biochemical events resulting in increased cell growthin vivoandin vitro1. The interaction of EGF with membrane receptors has been demonstrated in crude membrane preparations2, but no biochemical alteration of the membrane resulting from hormone binding has been detected. To clarify the molecular mechanisms regulating cell proliferation, specific biochemical reactions initiated by mitogens such as EGF need to be investigated in cell-free systems. As the human epidermoid carcinoma cell line A-431 has an extraordinarily high concentration of EGF receptors3,4(2–3 x 106receptors per cell), we have used a crude membrane preparation from these cells to look for an EGF-dependent alteration of membrane structure and/or function. We report here that (1) membranes may be prepared from A-431 cells that retain the ability to bind125I-labelled EGF in a specific manner, and (2) the binding of EGF to these membranesin vitroresults in a marked stimulation of the phosphorylation of endogenous proteins in the presence of [γ-32P]ATP.