Single-Molecule Super-Resolution Microscopy Reveals Heteromeric Complexes of MET and EGFR upon Ligand Activation
Single-Molecule Super-Resolution Microscopy Reveals Heteromeric Complexes of MET and EGFR upon Ligand Activation
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DOI：
10.3390/ijms21082803
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发表时间：
2020-04-01
影响因子：
5.6
通讯作者：
Heilemann, Mike
中科院分区：
文献类型：
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作者：
Harwardt, Marie-Lena I. E.;Schroeder, Mark S.;Heilemann, Mike
受体酪氨酸激酶（RTKs）协调细胞的运动和分化。不受管制的rtk可能促进癌症，是特定抑制剂的主要靶点。越来越多的证据表明，对抑制剂治疗的抗性涉及受体交叉相互作用，通过激活替代信号通路来绕过一种RTK的抑制。在这里，我们使用单分子超分辨率显微镜同时观察固定细胞和活细胞中HeLa和BT-20两种癌细胞系中的单个MET和表皮生长因子受体（EGFR）集群。我们在两种细胞类型中都发现了EGFR和MET的异聚受体簇，这是由配体激活促进的。活细胞中的单蛋白跟踪实验显示MET和EGFR对其同源和非同源配体的反应都是通过较慢的扩散。总之，我们首次提供了静态和动态证据，证明细胞膜上存在MET和EGFR的异聚簇，这与两种受体的相对表面表达水平相关。
Receptor tyrosine kinases (RTKs) orchestrate cell motility and differentiation. Deregulated RTKs may promote cancer and are prime targets for specific inhibitors. Increasing evidence indicates that resistance to inhibitor treatment involves receptor cross-interactions circumventing inhibition of one RTK by activating alternative signaling pathways. Here, we used single-molecule super-resolution microscopy to simultaneously visualize single MET and epidermal growth factor receptor (EGFR) clusters in two cancer cell lines, HeLa and BT-20, in fixed and living cells. We found heteromeric receptor clusters of EGFR and MET in both cell types, promoted by ligand activation. Single-protein tracking experiments in living cells revealed that both MET and EGFR respond to their cognate as well as non-cognate ligands by slower diffusion. In summary, for the first time, we present static as well as dynamic evidence of the presence of heteromeric clusters of MET and EGFR on the cell membrane that correlates with the relative surface expression levels of the two receptors.