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Sci Rep. 2016 Feb 15;6:21584. doi: 10.1038/srep21584.

Constructing modular and universal single molecule tension sensor using protein G to study mechano-sensitive receptors.

Wang X1,2,3, Rahil Z1,4, Li IT1,5, Chowdhury F2,6, Leckband DE4, Chemla YR1, Ha T1,2,7,8,9.

Author information

Department of Physics, Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Department of Physics and Astronomy, Iowa State University, Ames, Iowa 50011, USA.
Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Department of Chemistry, University of British Columbia Okanagan, Kelowna, BC, V1V1V7, Canada.
Mechanical Engineering and Energy Processes, Southern Illinois University, Carbondale, Illinois 62901, USA.
Howard Hughes Medical Institute, Baltimore, MD, 21218, USA.
Department of Biophysics &Biophysical Chemistry, Johns Hopkins University, Baltimore, MD 21205, USA.
Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD 21205, USA.


Recently a variety of molecular force sensors have been developed to study cellular forces acting through single mechano-sensitive receptors. A common strategy adopted is to attach ligand molecules on a surface through engineered molecular tethers which report cell-exerted tension on receptor-ligand bonds. This approach generally requires chemical conjugation of the ligand to the force reporting tether which can be time-consuming and labor-intensive. Moreover, ligand-tether conjugation can severely reduce the activity of protein ligands. To address this problem, we developed a Protein G (ProG)-based force sensor in which force-reporting tethers are conjugated to ProG instead of ligands. A recombinant ligand fused with IgG-Fc is conveniently assembled with the force sensor through ProG:Fc binding, therefore avoiding ligand conjugation and purification processes. Using this approach, we determined that molecular tension on E-cadherin is lower than dsDNA unzipping force (nominal value: 12 pN) during initial cadherin-mediated cell adhesion, followed by an escalation to forces higher than 43 pN (nominal value). This approach is highly modular and potentially universal as we demonstrate using two additional receptor-ligand interactions, P-selectin &PSGL-1 and Notch &DLL1.

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