Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell.
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 ~1 pN (10(-12) N), ~3 nm and ~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.
Woodruff School of Mechanical Engineering, Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology.
Yunfeng
Y
Chen
Coulter Department of Biomedical Engineering, Georgia Institute of Technology.
Baoyu
B
Liu
Charles Perkins Centre, The University of Sydney.
Lining
L
Ju
Woodruff School of Mechanical Engineering, Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology.
Jinsung
J
Hong
Institute of Biophysics, Laboratory of RNA Biology, Chinese Academy of Sciences; University of Chinese Academy of Sciences.
Qinghua
Q
Ji
School of Medicine and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University.
Wei
W
Chen
Woodruff School of Mechanical Engineering, Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology; cheng.zhu@bme.gatech.edu.
Cheng
C
Zhu
D020013:Calcium Signaling; D004912:Erythrocytes / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D055595:Mechanical Phenomena; D008856:Microscopy, Fluorescence / Q000379:methods*; D011485:Protein Binding; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D059010:Single-Cell Analysis / Q000379:methods*
D005456:Fluorescent Dyes; D008024:Ligands; D011948:Receptors, Antigen, T-Cell