Identification of a pivotal endocytosis motif in c-Met and selective modulation of HGF-dependent aggressiveness of cancer using the 16-mer endocytic peptide
Identification of a pivotal endocytosis motif in c-Met and selective modulation of HGF-dependent aggressiveness of cancer using the 16-mer endocytic peptide
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DOI：
10.1038/onc.2012.122
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发表时间：
2013-02-21
期刊：
影响因子：
8
通讯作者：
Park, Y. W.
中科院分区：
文献类型：
--
作者：
Cho, K-W;Park, J. H.;Park, Y. W.
由于c-Met在癌症的发展中具有重要作用，因此被认为是癌症治疗的有吸引力的靶点。虽然c-Met致癌特性的分子机制已被积极研究，c-Met内吞作用的调控元件及其对c-Met信号传导的影响仍不清楚。在这项研究中，我们确定了一个关键的内吞基序在c-Met和测试它的选择性调节HGF诱导的c-Met的反应。使用各种嵌合体与c-Met的胞质尾，我们能够证明，位于c-Met的C-末端的双亮氨酸基序的行为，以调节其内吞作用。合成肽Ant-3S由触角介导的蛋白质转导结构域（命名为Ant）和c-Met介导的16个氨基酸（命名为3S，跨越氨基酸1378至1393）组成，迅速移动到癌细胞中并破坏c-Met运输。重要的是，Ant-3S肽延长c-Met在膜上的保留时间显著降低磷酸化依赖性c-Met信号转导。此外，该肽有效地抑制HGF诱导的细胞生长、散射和迁移。已经研究了这些观察结果的潜在分子机制，并揭示了双亮氨酸基序与内吞机制（包括适应蛋白β和小窝蛋白-1）相互作用，以持续和增强信号转导。最后，Ant-3S肽特异性地阻断白细胞介素-2受体α亚基/3S嵌合蛋白的内化，但不阻断其他受体，包括Glut 4、Glut 8和转铁蛋白受体。这样的结果表明存在c-Met的选择性内吞组装。这也表明了使用本研究中鉴定的内吞基序进行c-Met特异性抗癌治疗的潜力。Oncogene（2013）32，1018-1029; doi：10.1038/onc.2012.122; 2012年4月23日在线发表
Since c-Met has an important role in the development of cancer, it is considered as an attractive target for cancer therapy. Although molecular mechanisms for oncogenic property of c-Met have been actively investigated, regulatory elements for c-Met endocytosis and its effect on c-Met signaling remain unclear. In this study, we identified a pivotal endocytic motif in c-Met and tested it for selective modulation of HGF-induced c-Met response. Using various chimeric constructs with the cytoplasmic tail of c-Met, we were able to demonstrate that a dileucine motif located in the C-terminus of c-Met acts to regulate its endocytosis. Synthetic peptide Ant-3S, consisting of antennapedia-derived protein transduction domain (designated as Ant) and c-Met-derived 16 amino-acids (designated as 3S, spanning amino-acids 1378 to 1393), rapidly moved into cancer cells and disrupted c-Met trafficking. Importantly, an extension of c-Met retention time on the membrane by Ant-3S peptide significantly decreased phosphorylation-dependent c-Met signal transduction. Additionally, the peptide effectively inhibited HGF-induced cell growth, scattering and migration. The underlying molecular mechanism for these observations has been investigated and revealed that the dileucine motif interacts with endocytic machinery, including adaptin beta and caveolin-1, for sustained and enhanced signal transduction. Finally, Ant-3S peptide specifically blocked internalization of interleukin-2 receptor alpha-subunit/3S chimeric protein, but not the other receptors, including Glut4, Glut8 and transferrin receptor. Such results indicate the presence of a selective endocytic assembly for c-Met. It also suggests a potential for c-Met-specific anti-cancer therapy using the identified endocytic motif in this study. Oncogene (2013) 32, 1018-1029; doi:10.1038/onc.2012.122; published online 23 April 2012