Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell
Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell
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DOI：
10.3791/52975
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发表时间：
2015-08-01
影响因子：
1.2
通讯作者：
Zhu, Cheng
中科院分区：
文献类型：
--
作者：
Chen, Yunfeng;Liu, Baoyu;Zhu, Cheng
膜受体-配体的相互作用介导了许多细胞功能。由这些分子相互作用触发的结合动力学和下游信号很可能受到发生结合和信号传递的机械环境的影响。最近的一项研究表明，机械力可以调节T细胞受体(TCR)对抗原的识别和触发。这是由我们开发的一项名为荧光生物膜力探针(FBFP)的新技术实现的，该技术将单分子力谱与荧光显微镜相结合。使用超软的人体红细胞作为敏感力传感器，采用高速摄像机和实时成像跟踪技术，fBFP在力、空间和时间分辨率上类似于1Pn(10(-12)N)，类似于3 nm，类似于0.5msec。有了fBFP，人们可以精确地测量力调节下单个受体-配体结合的动力学，并同时成像单个活细胞上结合触发的细胞内钙信号。这项新技术可用于研究其他细胞在机械调节下的膜受体-配体相互作用和信号转导。
Membrane receptor-ligand interactions mediate many cellular functions. Binding kinetics and downstream signaling triggered by these molecular interactions are likely affected by the mechanical environment in which binding and signaling take place. A recent study demonstrated that mechanical force can regulate antigen recognition by and triggering of the T-cell receptor (TCR). This was made possible by a new technology we developed and termed fluorescence biomembrane force probe (fBFP), which combines single-molecule force spectroscopy with fluorescence microscopy. Using an ultra-soft human red blood cell as the sensitive force sensor, a high-speed camera and real-time imaging tracking techniques, the fBFP is of similar to 1 pN (10(-12) N), similar to 3 nm and similar to 0.5 msec in force, spatial and temporal resolution. With the fBFP, one can precisely measure single receptor-ligand binding kinetics under force regulation and simultaneously image binding-triggered intracellular calcium signaling on a single live cell. This new technology can be used to study other membrane receptor-ligand interaction and signaling in other cells under mechanical regulation.