Quantum dot ligands provide new insights into erbB/HER receptor-mediated signal transduction
Quantum dot ligands provide new insights into erbB/HER receptor-mediated signal transduction
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DOI：
10.1038/nbt929
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发表时间：
2004-02-01
影响因子：
46.9
通讯作者：
Jovin, TM
中科院分区：
文献类型：
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作者：
Lidke, DS;Nagy, P;Jovin, TM
跨膜受体酪氨酸激酶（RTK）的erbB/HER家族介导细胞对表皮生长因子（EGF）及相关配体的反应。我们利用以下方法对活细胞中依赖RTK的信号传导的早期阶段进行了成像：（i）稳定表达与可见荧光蛋白（VFPs）融合的erbB1/2/3；（ii）带有表皮生长因子的荧光量子点（EGF - QD）；（iii）连续共聚焦激光扫描显微镜和流式细胞术。在此我们证明，EGF - QDs在表皮生长因子受体（erbB1）的结合和激活方面具有高度特异性和高效性，能迅速被内化到内涵体中，这些内涵体表现出活跃的运输和广泛的融合。结合在丝状伪足上表达的erbB1的EGF - QDs揭示了一种先前未报道的向细胞体逆行运输的机制。当erbB2 - 单体黄色荧光蛋白（mYFP）或erbB3 - 单体黄晶蛋白（mCitrine）与erbB1共表达时，EGF - QD和RTK - VFP的内吞速率和程度表明，在EGF刺激后，erbB2而非erbB3与erbB1异源二聚化，从而调节EGF诱导的信号传导。量子点 - 配体将在基础研究和生物技术发展中得到广泛应用。
The erbB/HER family of transmembrane receptor tyrosine kinases (RTKs) mediate cellular responses to epidermal growth factor (EGF) and related ligands. We have imaged the early stages of RTK-dependent signaling in living cells using: (i) stable expression of erbB1/2/3 fused with visible fluorescent proteins (VFPs), (ii) fluorescent quantum dots (QDs) bearing epidermal growth factor (EGF-QD) and (iii) continuous confocal laser scanning microscopy and flow cytometry. Here we demonstrate that EGF-QDs are highly specific and potent in the binding and activation of the EGF receptor (erbB1), being rapidly internalized into endosomes that exhibit active trafficking and extensive fusion. EGF-QDs bound to erbB1 expressed on filopodia revealed a previously unreported mechanism of retrograde transport to the cell body. When erbB2-monomeric yellow fluorescent protein (mYFP) or erbB3-monomeric Citrine (mCitrine) were coexpressed with erbB1, the rates and extent of endocytosis of EGF-QD and the RTK-VFP demonstrated that erbB2 but not erbB3 heterodimerizes with erbB1 after EGF stimulation, thereby modulating EGF-induced signaling. QD-ligands will find widespread use in basic research and biotechnological developments.