A General Approach for Generating Fluorescent Probes to Visualize Piconewton Forces at the Cell Surface
A General Approach for Generating Fluorescent Probes to Visualize Piconewton Forces at the Cell Surface
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DOI：
10.1021/jacs.5b11602
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发表时间：
2016-03-09
影响因子：
15
通讯作者：
Salaita, Khalid
中科院分区：
文献类型：
--
作者：
Chang, Yuan;Liu, Zheng;Salaita, Khalid
细胞与其细胞外基质（ECM）之间的机械力由数十种不同的受体介导。这些生物物理相互作用在从细胞发育到肿瘤进展的过程中起着重要作用。然而，映射各种受体配体对之间的张力的空间和时间动态仍然是一个重大的挑战。为了解决这个问题，我们已经开发了一种合成策略，以产生模块化的张力探针结合天然化学连接（NCL）反应与固相肽合成（SPPS）。原则上，这种方法实际上适应于任何适合于NCL的肽或表达的蛋白质。我们产生了一个小的张力探针库，显示不同的配体，柔性接头，和荧光报告，使整合素和钙粘蛋白张力的映射，并展示了长期（类似于3天）分子张力成像的第一个例子。这种方法提供了一个工具集，以更好地理解细胞生物学的基本机械转导事件。
Mechanical forces between cells and their extracellular matrix (ECM) are mediated by dozens of different receptors. These biophysical interactions play fundamental roles in processes ranging from cellular development to tumor progression. However, mapping the spatial and temporal dynamics of tension among various receptor ligand pairs remains a significant challenge. To address this issue, we have developed a synthetic strategy to generate modular tension probes combining the native chemical ligation (NCL) reaction with solid phase peptide synthesis (SPPS). In principle, this approach accommodates virtually any peptide or expressed protein amenable to NCL. We generated a small library of tension probes displaying different ligands, flexible linkers, and fluorescent reporters, enabling the mapping of integrin and cadherin tension, and demonstrating the first example of long-term (similar to 3 days) molecular tension imaging. This approach provides a toolset to better understand mechanotransduction events fundamental to cell biology.