Abstract：
Molecular forces are increasingly recognized as an important parameter to understand cellular signaling processes. In the recent years, evidence accumulated that also T-cells exert tensile forces via their T-cell receptor during the antigen recognition process. To measure such intercellular pulling forces, one can make use of the elastic properties of spider silk peptides, which act similar to Hookean springs: increased strain corresponds to increased stress applied to the peptide. Combined with Förster resonance energy transfer (FRET) to read out the strain, such peptides represent powerful and versatile nanoscopic force sensing tools. In this paper, we provide a detailed protocol how to synthesize a molecular force sensor for application in T-cell antigen recognition and hands-on guidelines on experiments and analysis of obtained single molecule FRET data.
摘要：
分子力越来越被认为是理解细胞信号传导过程的重要参数。近年来,积累的证据表明，在抗原识别过程中，T细胞也通过其T细胞受体施加拉力。为了测量这种细胞间拉力，可以利用蜘蛛丝肽的弹性特性，其作用类似于Hookean弹簧:增加的应变对应于施加到肽的增加的应力。结合福斯特共振能量转移(FRET)读出应变，这样的肽代表强大的和通用的纳米级力传感工具。在本文中,我们提供了一个详细的协议如何合成分子力传感器应用于T细胞抗原识别和实践指南的实验和分析获得的单分子FRET数据。
