A Mechanogenetic Toolkit for Interrogating Cell Signaling in Space and Time.
A Mechanogenetic Toolkit for Interrogating Cell Signaling in Space and Time.
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DOI：
10.1016/j.cell.2016.04.045
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发表时间：
2016-06-02
期刊：
影响因子：
64.5
通讯作者：
Jun YW
中科院分区：
文献类型：
--
作者：
Seo D;Southard KM;Kim JW;Lee HJ;Farlow J;Lee JU;Litt DB;Haas T;Alivisatos AP;Cheon J;Gartner ZJ;Jun YW
能够成像和扰动机械信号通路与精细的时空分辨率的工具一直难以捉摸，尽管它们在不同的细胞过程中的重要性。开发机械发生工具包（即，选择性和定量激活遗传编码的机械感受器）的挑战源于以下事实：许多机械激活的过程在空间和时间上是局部化的，但另外需要机械负载才能被激活。为了应对这一挑战，我们合成了磁等离子体纳米颗粒，可以成像，定位和机械加载具有高时空分辨率的靶蛋白。我们通过研究两种机械受体：Notch和E-cadherin的细胞表面活化来证明它们的实用性。通过测量单分子和单细胞水平的空间，化学，时间和机械输入的频谱的细胞反应，我们揭示了空间隔离和机械力如何合作，直接受体激活动力学。这种可推广的技术可用于控制和理解细胞信号传导中的各种机械敏感过程。
Tools capable of imaging and perturbing mechanical signaling pathways with fine spatiotemporal resolution have been elusive despite their importance in diverse cellular processes. The challenge in developing a mechanogenetic toolkit (i.e. selective and quantitative activation of genetically encoded mechanoreceptors) stems from the fact that many mechanically-activated processes are localized in space and time, yet additionally require mechanical loading to become activated. To address this challenge, we synthesized magnetoplasmonic nanoparticles that can image, localize, and mechanically load targeted proteins with high spatiotemporal resolution. We demonstrate their utility by investigating the cell surface activation of two mechanoreceptors: Notch and E-cadherin. By measuring cellular responses to a spectrum of spatial, chemical, temporal, and mechanical inputs at the single molecule and single cell level, we reveal how spatial segregation and mechanical force cooperate to direct receptor activation dynamics. This generalizable technique can be used to control and understand diverse mechanosensitive processes in cell signaling.