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Constructing modular and universal single molecule tension sensor using protein G to study mechano-sensitive
Xuefeng Wang1,2,3, Zainab Rahil1,4, Isaac T S Li1,5
1Department of Physics, Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Scientific Reports
|February 16, 2016
Summary
This study introduces a novel Protein G-based force sensor for measuring cellular forces on single receptors. This method simplifies measurements by avoiding direct ligand conjugation, revealing new insights into cell adhesion forces.
Area of Science:
- Biophysics
- Cell Biology
- Biochemistry
Background:
- Molecular force sensors are crucial for studying cellular forces on mechano-sensitive receptors.
- Current methods often involve complex, labor-intensive ligand conjugation to force-reporting tethers, potentially reducing ligand activity.
Purpose of the Study:
- To develop a simplified and modular molecular force sensor system.
- To overcome limitations of direct ligand conjugation in previous force sensing techniques.
Main Methods:
- Developed a Protein G (ProG)-based force sensor where tethers attach to ProG, not ligands.
- Utilized ProG:Fc binding for convenient assembly of recombinant ligands fused with IgG-Fc.
- Applied the sensor to measure forces in E-cadherin-mediated cell adhesion, P-selectin &PSGL-1, and Notch &DLL1 interactions.
Main Results:
- Demonstrated that molecular tension on E-cadherin is initially lower than the dsDNA unzipping force (12 pN).
- Observed an escalation of tension on E-cadherin to forces exceeding 43 pN during cell adhesion.
- Validated the modularity and universality of the ProG-based sensor across multiple receptor-ligand systems.
Conclusions:
- The ProG-based force sensor offers a streamlined, efficient, and versatile approach for measuring molecular forces in cellular processes.
- This method avoids ligand conjugation, preserving ligand activity and simplifying experimental workflows.
- The sensor provides valuable quantitative data on forces involved in key cell adhesion and signaling pathways.
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