Interaction of the extracellular domain of the epidermal growth factor receptor with gangliosides
Interaction of the extracellular domain of the epidermal growth factor receptor with gangliosides
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DOI：
10.1074/jbc.m111669200
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发表时间：
2002-03-22
影响因子：
4.8
通讯作者：
Bremer, EG
中科院分区：
文献类型：
--
作者：
Miljan, EA;Meuillet, EJ;Bremer, EG
神经节苷脂GM 3在多种细胞系中抑制表皮生长因子（EGF）依赖性细胞增殖。在体外和体内，这种鞘糖脂抑制EGF受体（EGFR）的激酶活性。此外，含有EGFR的膜制剂可以结合到GM 3包被的表面。这些数据表明GM 3可能与EGFR直接相互作用。在这项研究中，神经节苷脂与EGFR的胞外结构域（ECD）的相互作用进行了研究。从昆虫细胞中纯化的人重组ECD直接与神经节苷脂GM 3结合。神经节苷脂相互作用的网站似乎是不同的EGF结合位点。与先前关于特定神经节苷脂对EGFR激酶活性的影响的报道一致，ECD优先与GM 3相互作用。其他神经节苷脂的相对结合力大小顺序为：GM 3> GM 2，GD 3，GM 4> GM 1，GD 1a，GD 1b，GT 1b，GD 2，GQ 1b>乳糖神经酰胺。这些数据表明，NeuAc-乳糖是必不可少的结合和任何糖取代减少结合。与可溶性ECD结合神经节苷脂的特异性一致，GM 3特异性抑制EGFR自磷酸化。EGFR ECD上神经节苷脂相互作用位点的鉴定与内源性GM 3可能作为EGFR活性的直接调节剂起作用的假设一致。
Ganglioside GM3 inhibits epidermal growth factor (EGF)-dependent cell proliferation in a variety of cell lines. Both in vitro and in vivo, this glycosphingolipid inhibits the kinase activity of the EGF receptor (EGFR). Furthermore, membrane preparations containing EGFR can bind to GM3-coated surfaces. These data suggest that GM3 may interact directly with the EGFR. In this study, the interaction of gangliosides with the extracellular domain (ECD) of the EGFR was investigated. The purified human recombinant ECD from insect cells bound directly to ganglioside GM3. The ganglioside interaction site appears to be distinct from the EGF-binding site. In agreement with previous reports on the effects of specific gangliosides on EGFR kinase activity, the ECD preferentially interacted with GM3. The order of relative binding of other gangliosides investigated was as follows: GM3 much greater than GM2, GD3, GM4 > GM1, GD1a, GD1b, GT1b, GD2, GQ1b > lactosylceramide. These data suggest that NeuAc-lactose is essential for binding and that any sugar substitution reduces binding. In agreement with the specificity of soluble ECD binding to gangliosides, GM3 specifically inhibited EGFR autophosphorylation. Identification of a ganglioside interaction site on the ECD of the EGFR is consistent with the hypothesis that endogenous GM3 may function as a direct modulator of EGFR activity.