摘要: 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.
-
期刊:
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
-
分类:
生物学
>>
生物物理学
-
引用:
ChinaXiv:201605.01354
(或此版本
ChinaXiv:201605.01354V1)
DOI:10.12074/201605.01354V1
CSTR:32003.36.ChinaXiv.201605.01354.V1
- 推荐引用方式:
Chen, Yunfeng,Hong, Jinsung,Zhu, Cheng,Liu, Baoyu,Ju, Lining,Ji, Qinghua,Ji, Qinghua,Chen, Wei,Chen, Wei.(2016).Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell.JOVE-JOURNAL OF VISUALIZED EXPERIMENTS.[ChinaXiv:201605.01354]
(点此复制)